Novel ECL Method for the Determination of Skatole in Porcine Adipose Tissue.

A new method for the determination of skatole present in porcine adipose tissue samples utilizing the electrochemiluminescence of skatole is presented. It has been observed that oxygen radicals produced at a high cathodic voltage can react with oxidized skatole to create an excited intermediate molecule that then relaxes, generating peak photon emission at around 480 nm. A strong electrochemiluminescence or electrogenerated chemiluminescence (ECL) signal using boron-doped diamond (BDD) electrodes was observed optimally when a reduction potential of -1.8 V was applied, held for 40 s, before holding an oxidation potential of 0.8 V for 10 s. Using this principle, a calibration curve using known concentrations of skatole showed good linearity (range 0.025-2 µM) and a very low detection limit (LOD, 0.7 nM). A method that demonstrates for the first time an approach that utilizes this ECL reaction, and has the potential to be developed into an analytical device for use in the slaughterhouse, has been developed. This was achieved by extracting skatole out of the porcine adipose tissues into acetonitrile - giving an extraction efficiency of 67.6%. This method was then validated by analyzing the skatole content of 33 pig fat samples that had been previously tested using a standard technique, high-performance liquid chromatography (HPLC), containing a range of concentrations (0.02-2.58 µg/g). This ECL method exhibited excellent reliability and correlation with HPLC, giving a R2 coefficient of 0.911, thus demonstrating the potential for this method to be developed for an on-line skatole detector.

Effect of surgical or immune castration on postprandial nutrient profiles in male pigs.

To avoid boar taint before slaughtering, late castration by immunisation against gonadotrophin releasing hormone or immunocastration has been developed. The current study aimed at determining whether differences in feed efficiency between castrated male (CM), immune castrated (IM) and entire male pigs (EM) can be explained by differences in nutrient plasma profiles after a meal. In this study, 24 male pigs (n = 8/type) were enrolled between 14 to 19 weeks of age. Entire and IM pigs ate less and were more feed efficient than CM pigs (p < 0.05). The postprandial plasma profiles of glucose, insulin, urea and amino acids (AA) were determined before (d -6), just after (d 8) and well after (d 16) the decrease in testicular hormones in IM pigs. For each test day, pigs were fasted overnight and subsequently fed a small meal (400 g). On d -6, postprandial profiles of plasma glucose of IM pigs did not differ from the two other types of pigs. On d 8, EM pigs had a greater average plasma glucose concentration than IM and CM pigs (p < 0.05) but the profiles did not differ. On d 16, the differences between profiles of glucose suggest a lower clearance of glucose in EM compared with IM and CM pigs. Plasma insulin did not significantly differ between the three types. Plasma urea profiles did not differ between CM, EM and IC pigs on d -6 whereas CM pigs showed higher plasma urea concentrations than EM and IM thereafter (p < 0.01). Among AA, Lys plasma concentrations were greater in CM than in EM and IM pigs on d -6, d 8 and d 16 (p < 0.05), whereas on d 16 plasma Hypro concentrations were lower in CM than in EM and IM pigs (p < 0.05). The finding that plasma glucose profiles were modified by immunocastration much faster and earlier than urea and AA profiles, suggest that the decrease in testicular hormones impacted energy metabolism more rapidly than protein metabolism.

Plasma cortisol and faecal cortisol metabolites concentrations in stereotypic and non-stereotypic horses: do stereotypic horses cope better with poor environmental conditions?

BACKGROUND: Stereotypic behaviours, i.e. repetitive behaviours induced by frustration, repeated attempts to cope and/or brain dysfunction, are intriguing as they occur in a variety of domestic and captive species without any clear adaptive function. Among the different hypotheses, the coping hypothesis predicts that stereotypic behaviours provide a way for animals in unfavourable environmental conditions to adjust. As such, they are expected to have a lower physiological stress level (glucocorticoids) than non-stereotypic animals. Attempts to link stereotypic behaviours with glucocorticoids however have yielded contradictory results. Here we investigated correlates of oral and motor stereotypic behaviours and glucocorticoid levels in two large samples of domestic horses (NStudy1 = 55, NStudy2 = 58), kept in sub-optimal conditions (e.g. confinement, social isolation), and already known to experience poor welfare states. Each horse was observed in its box using focal sampling (study 1) and instantaneous scan sampling (study 2). Plasma samples (collected in study 1) but also non-invasive faecal samples (collected in both studies) were retrieved in order to assess cortisol levels. RESULTS: Results showed that 1) plasma cortisol and faecal cortisol metabolites concentrations did not differ between horses displaying stereotypic behaviours and non-stereotypic horses and 2) both oral and motor stereotypic behaviour levels did not predict plasma cortisol or faecal cortisol metabolites concentrations. CONCLUSIONS: Cortisol measures, collected in two large samples of horses using both plasma sampling as well as faecal sampling (the latter method minimizing bias due to a non-invasive sampling procedure), therefore do not indicate that stereotypic horses cope better, at least in terms of adrenocortical activity.

Combining sheep and beef cattle in a pasture-based system minorly influenced muscle and fat colour and dorsal fat firmness but increased fat skatole content in lambs.

A recent long-term system-level experiment, using grassland-based organic systems established as separate farmlets, showed that combining sheep and beef cattle within a mixed (MIX) system promoted self-sufficient production of grass-fed meat and improved economic and environmental performance in the sheep enterprise compared to a sheep-only (SH) system. In the present study, we used all male lambs produced over the first year in this experiment to evaluate the effect of this farming practice on some lamb carcass and meat sensory quality traits. Lambing took place at the end of February and lambs were pasture-fed from 1 month of age on average until slaughter, unless they were not ready for slaughter at 3-4 weeks before the start of the mating period in October, in which case they were finished indoors on a concentrate-based diet. All MIX lambs (n = 33) were finished at pasture. One SH lamb (out of a total of 85) had to be finished indoors with a concentrate-based diet. MIX lambs had a higher growth rate than SH lambs (P < 0.001), resulting in a lower lamb age at slaughter in MIX than in SH (150 vs 173 days, P < 0.001). Carcass weight and degree of fatness, kidney fat weight and dorsal fat thickness did not differ between systems. Kidney fat skatole concentration was higher in MIX than in SH (median value reaching 0.15 µg/g liquid fat and 0.11 µg/g liquid fat in MIX and SH, respectively, P < 0.05). This was most likely due to MIX lambs being slaughtered at a younger age and therefore eating younger grass. There were minor differences between MIX and SH lambs in dorsal fat firmness and in colour coordinates of longissimus thoracis et lumborum muscle. We conclude that while combining sheep and beef cattle within a mixed system offers advantages in terms of self-sufficient production of grass-fed meat and system economic and environmental performance, it could come at a cost to lamb flavour.

Change in fat skatole and indole content in lambs switched from a concentrate-based diet indoors to alfalfa grazing for various durations before slaughter.

Meat from lambs fattened on alfalfa is at risk of excessive pastoral flavours due to high levels of in-fat volatile indolic compounds (especially, skatole). Skatole has also been identified as a potential marker of interest for authenticating pasture-fed lamb meat. Here, we investigated the change in skatole and indole concentrations in kidney fat from lambs switched from an indoor-fed concentrate based diet to outdoor alfalfa grazing for various durations (0, 21, 42, 63 days) before slaughter. The study used a total of 219 lambs over 3 consecutive years. Kidney-fat skatole and indole concentrations increased from as early as 21 days on alfalfa, and then reached a plateau. Similarly, the proportion of lambs that had a kidney fat-skatole concentration above 0.15 µg/g liquid fat, a value that has been established as a sensory rejection threshold for pork, increased significantly from as early as 21 days on alfalfa and then reached a plateau. This value was reached or exceeded in a significant proportion of lambs fattened on alfalfa pastures (45.1%). However, skatole was not detected in kidney fat from 20 out of 164 alfalfa-fattened lambs (i.e., 12.2%) but was detected in 15 out of 55 concentrate-fed lambs (i.e., 27.3%). We thus conclude that while skatole content in kidney fat can inform on dietary changes made shortly before slaughter, it does not have the discrimination power needed to reliably authenticate pasture-fed lamb meat, let alone duration of finishing on pasture.

Is a short concentrate-finishing period in lambs raised on alfalfa pasture effective at reducing fat indoles contents and lightening meat colour?

Pasture-feeding in lambs has been associated with dark-coloured meat and high meat contents of volatile indoles (skatole and indole), which may be responsible for sensory defects. The risk of off-flavours is even higher when lambs graze alfalfa, because it is particularly rich in rapidly degradable proteins. Here, we investigated whether a short concentrate stall-finishing period in lambs that previously grazed alfalfa influences meat sensory quality traits. We compared three feeding treatments, using three groups of 10 male Romane lambs: grazing alfalfa (A), stall-feeding with concentrate and straw (S), and stall-finishing with concentrate and straw for 21 days after previously grazing alfalfa (AS). During stall-feeding, the concentrate was given at a level adjusted to achieve a similar growth pattern in all treatment groups, and the length of the trough was sufficient for all lambs to have access to the feeds at the same time. Lamb live weight and age at slaughter, and carcass conformation and fatness were similar for all treatment groups. Perirenal fat skatole content was lower in AS and S lambs than in A lambs (P < 0.001 and P < 0.05, respectively), and not significantly different between AS and S lambs. Perirenal fat indole and dorsal fat skatole concentrations were lower in AS lambs than in A lambs (P < 0.05 and P < 0.005, respectively), the other pairwise comparisons being not significant. There was no treatment effect on dorsal fat indole content. Longissimus et lumborum muscle colour coordinates differed between pasture-fed and stall-fed lambs, with a lower lightness (P < 0.005) and greater redness and hue angle (P < 0.001 and P < 0.01, respectively) in pasture-fed lambs, but muscle colour coordinates did not differ between A and AS lambs. A 21-d concentrate-based stall-finishing period in lambs previously raised on alfalfa pasture therefore markedly reduced fat indoles concentrations compared to lambs grazed on alfalfa until slaughter but did not change meat colour.

Evaluation of the potential benefits of iron supplementation in organic pig farming.

Background: Iron from the stock acquired during foetal life and the ingestion of milk is not sufficient to cover the needs of the piglets during their first weeks of life. In organic farming, systematic supplementation with iron is problematic due to a strong limitation in pharmaceutic treatments. Methods: Erythroid parameters around weaning were measured in piglets from organic outdoor and indoor farms, and related to indicators of the inflammatory status. Blood samples were collected from 28.9±2.6 piglets/herd at 42.0±3.2 days of age and 11.9±3.0 kg live weight (mean ± SD) in 21 farms from the west part of France. Among the 11 outdoor farms, only one had supplemented piglets with 200 mg iron while among the 10 indoor farms, only one had not supplemented piglets, one had supplemented them with 100 mg, 8 with 200 mg and one with 400 mg. Results: Compared to outdoor piglets without supplementation, piglets kept indoors and receiving 200 mg iron had lower haemoglobin concentration (105 vs 118±2 g/l, mean ± SE) and red blood cell volume (56 vs 60±1 fl) (P<0.005). The reduction in haemoglobin concentration and red blood cell volume was more pronounced in indoor piglets supplemented with 100 mg of iron and even more when they had not received iron. The plasma concentration of haptoglobin was lower in outdoor than in indoor piglets (0.51±0.06 vs 0.78±0.09 g/l) whereas no effect of housing was observed for markers of oxidative stress (dROM, BAP). In the 14 farms where sow parity was known, the haemoglobin concentration was lower in piglets from primiparous than from multiparous sows (109 versus 114±2 g/l, P < 0.001). Conclusion: With the exception of soils where the content of bioavailable iron is very low, piglets from outdoor farms do not require iron supplementation, unlike those raised indoors.

Improving maternal welfare during gestation has positive outcomes on neonatal survival and modulates offspring immune response in pigs.

Improving the housing of pregnant sows by giving them more space and access to deep straw had positive effects on their welfare, influenced their maternal behavior and improved the survival of their offspring. The present study aimed at determining whether these effects were actually due to environmental enrichment and whether the provision of straw pellets and wood can partly mimic the effects of straw bedding during gestation. Three graded levels of enrichment were used, that were, collective conventional pens on slatted floor (C, n = 26), the same pens with manipulable wood materials and distribution of straw pellets after the meals (CE, n = 30), and larger pens on deep straw litter (E, n = 27). Sows were then housed in identical farrowing crates from 105 days of gestation until weaning. Decreased stereotypies, blood neutrophils, and salivary cortisol, and increased behavioral investigation indicated that health and welfare of sows during gestation were improved in the E environment compared with the C environment. The CE sows responded as C or E sows depending on the trait. Piglet mortality rate in the first 12 h after birth was lower in E and CE litters than in C litters, but enrichment level during gestation had only small effects on lactating sow behavior and milk composition postpartum. On days 2 and 3 of lactation, E sows interrupted less often their nursing sequences than C and CE sows. On day 2, milk from both E and CE sows contained more minerals than that from C sows. In one-day-old piglets, the expression levels of genes encoding toll-like receptors (TLR2, TLR4) and cytokines (interleukin-1, -6 and -10) in whole blood after 20-h culture, were greater in E piglets than in CE or C piglets. In conclusion, housing sows in an enriched environment during gestation improved early neonatal survival, probably via moderate and cumulative positive effects on sow behavior, milk composition, and offspring innate immune response. The gradation in the effects observed in C, CE and E housing environment reinforced the hypothesis of a causal relationship between maternal environmental enrichment, sow welfare and postnatal piglet traits.

Correlation Networks Provide New Insights into the Architecture of Testicular Steroid Pathways in Pigs.

Steroid metabolism is a fundamental process in the porcine testis to provide testosterone but also estrogens and androstenone, which are essential for the physiology of the boar. This study concerns boars at an early stage of puberty. Using a RT-qPCR approach, we showed that the transcriptional activities of several genes providing key enzymes involved in this metabolism (such as CYP11A1) are correlated. Surprisingly, HSD17B3, a key gene for testosterone production, was absent from this group. An additional weighted gene co-expression network analysis was performed on two large sets of mRNA-seq to identify co-expression modules. Of these modules, two containing either CYP11A1 or HSD17B3 were further analyzed. This comprehensive correlation meta-analysis identified a group of 85 genes with CYP11A1 as hub gene, but did not allow the characterization of a robust correlation network around HSD17B3. As the CYP11A1-group includes most of the genes involved in steroid synthesis pathways (including LHCGR encoding for the LH receptor), it may control the synthesis of most of the testicular steroids. The independent expression of HSD17B3 probably allows part of the production of testosterone to escape this control. This CYP11A1-group contained also INSL3 and AGT genes encoding a peptide hormone and an angiotensin peptide precursor, respectively.

Pre-emptive meloxicam for postoperative analgesia in piglets undergoing surgical castration.

OBJECTIVE: To investigate the effect of preoperative meloxicam administration on postoperative stress and pain induced by surgical castration in piglets. STUDY DESIGN: Prospective, blinded, randomized clinical trial. ANIMALS: One hundred and eighty male piglets of <1 week of age. METHODS: Castration was performed on 150 piglets which had received either an intramuscular injection of 0.4 mg kg(-1) meloxicam or a placebo 10-30 minutes before the procedure. Blood cortisol and ACTH concentrations were determined at 30 minutes post-castration and haptoglobin was measured at 24 hours post-castration. Presence or absence of foreleg movements, hind leg movements, urine or faeces emission, tremors or other body movements were recorded during the castration procedure. Scores for presence or absence of prostration, tremors, tail movements and isolation were recorded at 30 minutes, and at 1, 2, 4 and 24 hours post-castration and combined in a global behaviour score (GBS). Blood samples were taken from a further 30 piglets which did not undergo castration. RESULTS: Mean blood cortisol and ACTH concentrations at 30 minutes post-castration were both significantly lower in the meloxicam group than in the placebo group (p < or = 0.01). The mean haptoglobin concentration at 24 hours was not significantly reduced (p = 0.178). The distribution of the GBS during castration was similar in both groups. There were significant differences in the GBS after castration at both 2 and 4 hours post-castration with a greater proportion of piglets in the meloxicam group showing no behavioural alterations (82.7%versus 68.0% at both time points). The score distribution was similar in both groups at 30 minutes, 1 and 24 hours after castration. CONCLUSION AND CLINICAL RELEVANCE: This study suggests that pre-emptive administration of meloxicam is able to produce some postoperative analgesia after surgical castration of young piglets.

Welfare Aspects of Raising Entire Male Pigs and Immunocastrates.

For a long time, scientists assumed that newborns have a severely limited sense of pain (if any). However, this assumption is wrong and led to a "start of the exit" from piglet surgical castration. Some of the currently discussed or already implemented alternatives such as general or local anaesthesia during surgical castration raise additional welfare concerns as well as legal problems and/or are hardly applicable. The favoured long-term, welfare-friendly "gold standard" is to raise entire male pigs (EM). However, this may also impose certain welfare problems under the current conventional housing and management conditions. The specific types of behaviour displayed by EM such as mounting and aggressive behaviours but also increased exploration, which are partially linked to sexual maturation, increase the risk for injuries. The current status of knowledge (scientific literature and farmer experiences) on housing of EM suggests that environmental enrichment, space, group-stability, social constellation, feeding (diet and feeder space), health and climate control are critical factors to be considered for future housing systems. From an animal welfare point of view, an intermediate variant to be favoured to reduce problematic behaviour could be to slaughter EM before reaching puberty or to immunize boars early on to suppress testicular function. Immunization against endogenous GnRH can reduce EM-specific problems after the 2nd vaccination.

Evidence of Pain, Stress, and Fear of Humans During Tail Docking and the Next Four Weeks in Piglets (Sus scrofa domesticus).

Tail docking is widely performed in pig farms to prevent tail biting. We investigated the consequences of this practice on behavioral indicators of pain and stress, and on the human-piglet relationship during lactation. Within 19 litters, piglets (1-3 days of age) were submitted on day 0 (D0) to docking with a cautery iron (D), sham-docking (S), or no docking (U). Piglets from the D and S groups were observed during the procedure (body movements and vocalizations) and just after, in isolation, during 20 s for body, tail and ear postures as well as ear movements. Piglets from the three treatments were observed in their home pen after docking on D0 and D3 afternoon for body posture, tail posture and movements. Piglets from the D and U groups were observed on D6, D12, D19, and D26 in their home pen for oral behavior, body, and tail posture. Tail damage and tear staining were scored on D5, D11, D18, and D25. A 5-min motionless human test was performed on D14. During the procedure, D piglets screamed more and with a higher intensity (P < 0.05) than S piglets (n = 48-50). Just after docking, D piglets held their ears in a posture perpendicular to the head-tail axis and changed their ear posture more often (P < 0.05). Between D6 and D26, D piglets kept their tail immobile (P < 0.001) and in a horizontal position (P < 0.01) more often than U piglets (n = 45-47). Between D11 and D25, U piglets had higher scores for tail damage and damage freshness than D piglets (0.09 < P < 0.02) whereas tear-stain score was similar. In the human test, D piglets interacted later with an unfamiliar human than U piglets (P = 0.01, n = 18/group). Present data indicate signs of acute pain and stress in piglets due to docking during the procedure itself and adverse consequences throughout lactation thereafter, including on their relationship with humans. On the other hand, the presence of tail lesions shows that undocked piglets are subject to more tail biting, even before weaning.

Chronic intermittent stress exposure and access to grass silage interact differently in their effect on behaviour, gastric health and stress physiology of entire or castrated male growing-finishing pigs.

Entire male pigs display more aggressive and sexual behaviour. This might cause a condition of chronic stress and impair their welfare. In order to assess chronic stress in entire and castrated male pigs, as well as effects of providing grass silage as occupational and feed material on behaviour and health, we carried out a 2 × 2 × 2-factorial experiment with 147 growing-finishing pigs. Factors investigated were castration (entire/castrated), chronic intermittent social stress exposure (yes/no) and access to grass silage (yes/no), as well as their interactions. The stress exposure treatment consisted of repeated short-term confrontations and separations. We recorded different behavioural variables, circadian rhythm of salivary cortisol, response to an ACTH (adrenocorticotropic hormone) challenge test, pathological changes in the gastric mucosa and morphology of the intestinal epithelium. Stress exposure caused a decrease in posture changes and head knocks/bites in the home pen. Reference indicators affected by stress exposure did not differ between entire and castrated male pigs, indicating that there is no permanently increased baseline level of stress in entire male pigs. However, entire males responded more pronouncedly to the stress exposure compared to castrated males in terms of posture changes and play behaviour. Pigs provided with grass silage showed more play behaviour and less manipulative behaviours than pigs not receiving grass silage. Stress treated pigs had more hyperkeratosis in the gastric mucosa and gastric ulcers, while offering grass silage reduced such changes. In conclusion, our results indicate that the increased behavioural stress response of entire male pigs might require some adaptations in housing and management of entire male pigs. Gastric ulceration scoring turned out to be a potential post mortem indicator for chronic stress. Finally, providing roughages like grass silage could be a means to positively affect behaviour and gastric health in pigs.

Heart rate variability as a measure of autonomic regulation of cardiac activity for assessing stress and welfare in farm animals -- a review.

Measurement of heart rate variability (HRV) is a non-invasive technique that can be used to investigate the functioning of the autonomic nervous system, especially the balance between sympathetic and vagal activity. It has been proven to be very useful in humans for both research and clinical studies concerned with cardiovascular diseases, diabetic autonomic dysfunction, hypertension and psychiatric and psychological disorders. Over the past decade, HRV has been used increasingly in animal research to analyse changes in sympathovagal balance related to diseases, psychological and environmental stressors or individual characteristics such as temperament and coping strategies. This paper discusses current and past HRV research in farm animals. First, it describes how cardiac activity is regulated and the relationships between HRV, sympathovagal balance and stress and animal welfare. Then it proceeds to outline the types of equipment and methodological approaches that have been adapted and developed to measure inter-beats intervals (IBI) and estimate HRV in farm animals. Finally, it discusses experiments and conclusions derived from the measurement of HRV in pigs, cattle, horses, sheep, goats and poultry. Emphasis has been placed on deriving recommendations for future research investigating HRV, including approaches for measuring and analysing IBI data. Data from earlier research demonstrate that HRV is a promising approach for evaluating stress and emotional states in animals. It has the potential to contribute much to our understanding and assessment of the underlying neurophysiological processes of stress responses and different welfare states in farm animals.

Exploration of steroidogenesis-related genes in testes, ovaries, adrenals, liver and adipose tissue in pigs.

To explore the metabolism of steroids in the pig species, a qualitative PCR analysis was performed for the main transcript of 27 genes involved in steroid metabolism. We compared samples of testes, adipose tissue and liver from immature and peripubertal males, adrenal cortex from peripubertal males, ovaries from cyclic females and adipose tissue from peripubertal females. Some genes were shown to have a tissue-specific expression. Two of them were expressed only in testes, ovaries and adrenals: CYP11A1 and CYP11B. The CYP21 and HSD17B3 genes, were expressed respectively only in adrenals and only in testes. Very few differences were observed between transcriptional patterns of peripubertal testes and adrenal glands as well as between male and female fat tissues. However, the expression of genes involved in the sulfonation of steroids was higher in testes than in adrenals from males. Main differences between ovaries and testes were observed for HSD17B1/2/3, AKR1C-pig6 and sulfotransferase genes (SULT2A1/SULT2B1). The present study shows that the SRD5A2 and CYP21 genes were not involved in the testicular biosynthesis of androstenone. It also shows that porcine adrenal glands produce essentially corticosteroids and that fat tissue is unable to produce de novo steroids.

Dietary protein intake during the oestrous cycle does not alter the ovulation rate in gilts.

The effect of protein intake on ovulation rate was investigated in cross-bred gilts. On Day 14 of the third oestrous cycle, luteolysis was induced by injection of an analogue of prostaglandin F(2alpha). The ovulation rate was recorded when gilts were killed on Day 27. In the first experiment, nutritional treatment was applied from Day 14 to Day 27. Gilts were fed diets providing the same amount of digestible energy, but containing either a low, medium or high amount of dietary protein. Protein restriction linearly decreased plasma concentrations of urea (P < 0.001) and had no effect on plasma concentrations of insulin-like growth factor-I and leptin measured at Day 27 (P > 0.1). Protein restriction did not influence ovulation rate (mean (+/- s.e.m.) 17.0 +/- 0.4; P > 0.1). In the second experiment, the nutritional treatment was applied from Day 5 until Day 27. Gilts received the same amount of digestible energy and either a low or a high amount of protein, as in Experiment 1. A third group of gilts received the high amount of protein and also more digestible energy. The ovulation rate did not differ between the three groups of gilts. These data indicate no effect of short-term dietary protein restriction on ovulation rate in cyclic gilts.

Pathways and genes involved in steroid hormone metabolism in male pigs: a review and update.

This paper reviews state-of-the-art knowledge on steroid biosynthesis pathways in the pig and provides an updated characterization of the porcine genes involved in these pathways with particular focus on androgens, estrogens, and 16-androstenes. At least 21 different enzymes appear to be involved in these pathways in porcine tissues together with at least five cofactors. Until now, data on several porcine genes were scarce or confusing. We characterized the complete genomic and transcript sequences of the single porcine CYP11B gene. We analyzed the porcine AKR1 gene cluster and identified four AKR1C, one AKR1C like genes and one AKR1E2 gene. We provide evidence that porcine AKR1C genes are not orthologous to human AKR1C. A new nomenclature is thus needed for this gene family in the pig. Thirty-two genes are now described: transcript (30+2 characterized in this study) and genomic (complete: 18+1 and partial: 12+1) sequences are identified. However, despite increasing knowledge on steroid metabolism in the pig, there is still no explanation of why porcine testes can produce androstenone and epiandrosterone, but not dihydrotestosterone (DHT), which is also a reduced steroid.

Stress modulates instrumental learning performances in horses (Equus caballus) in interaction with temperament.

The present study investigates how the temperament of the animal affects the influence of acute stress on the acquisition and reacquisition processes of a learning task. After temperament was assessed, horses were subjected to a stressor before or after the acquisition session of an instrumental task. Eight days later, horses were subjected to a reacquisition session without any stressor. Stress before acquisition tended to enhance the number of successes at the beginning of the acquisition session. Eight days later, during the reacquisition session, contrary to non-stressed animals, horses stressed after acquisition, and, to a lesser extent, horses stressed before acquisition, did not improve their performance between acquisition and reacquisition sessions. Temperament influenced learning performances in stressed horses only. Particularly, locomotor activity improved performances whereas fearfulness impaired them under stressful conditions. Results suggest that direct exposure to a stressor tended to increase acquisition performances, whereas a state of stress induced by the memory of a stressor, because it has been previously associated with the learning context, impaired reacquisition performances. The negative effect of a state of stress on reacquisition performances appeared to be stronger when exposure to the stressor occurred after rather than before the acquisition session. Temperament had an impact on both acquisition and reacquisition processes, but under stressful conditions only. These results suggest that stress is necessary to reveal the influence of temperament on cognitive performances.

Maternal stress during late gestation has moderate but long-lasting effects on the immune system of the piglets.

Events acting prenatally on developing foetuses are important determinants for disorders later in life. Prenatal stress (PNS) is one of these events. The purpose of this study was to determine the consequences of a repeated social stress applied during late gestation of the pregnant gilt on the immune system and hypothalamo-pituitary-adrenal (HPA) axis activity of the piglets from birth to two months of age. Pregnant gilts were submitted to repeated social stress which was induced by housing unfamiliar gilts in pairs modified twice a week during 4 weeks between days 77 and 105 of gestation (S group, n=18). Control gilts were housed in stable pairs during the same period (C group, n=18). Blood cortisol, haptoglobin and IgG levels, immune cell counts, mitogen-induced whole-blood proliferation and TNF-alpha production were evaluated in piglets at 4 days of age (D4), before and after weaning (D26 and 28) and before and after relocation to a new building (D60 and 62). We found that PNS did not affect growth rate of the progeny. It decreased the relative weight of adrenal glands on D4 (P<0.05) but plasma cortisol levels were similar in both groups at all ages. IgG levels in colostrum and in the serum of piglets were not affected. PNS decreased the total numbers of white blood cells, lymphocytes and granulocytes from D26 to D60 (P<0.05), the CD4(+)/CD8(+) T cell ratio on D4 (P<0.05), and LPS induced-TNF-alpha production on D60 (P<0.05). PNS increased the ConA-induced lymphocyte proliferation on D4 and D60 and the PWM-induced proliferation on D60 (P<0.05). Our results demonstrate that a repeated social stress applied to pregnant sows during late gestation can induce long-lasting effects on several parameters of the immune function of the offspring. These effects are not due to modifications of the HPA axis activity and may impair the abilities of the piglets to efficiently react against infections during the suckling period and around weaning.

Comparative effects of a prenatal stress occurring during early or late gestation on pig immune response.

The aim of this work was to investigate the immune effects of prenatal stress (PNS) in pigs, when maternal stressor is applied during early or late gestation. In two separate experiments, gilts were submitted to a social-stress routine between either days 24 and 48 (stress (S) group, n=8 vs. control (C) group, n=6) or days 79 and 103 of gestation (S group, n=10 vs. C group, n=7). During the first month of life of the piglets, their cortisol levels were assessed in basal state and after stressful events (castration, new environment, and weaning). Piglets were immunized against ovalbumin (OVA) at 7 (D7) and 19 (D19) days of age. Lymphocyte proliferation in response to OVA and mitogens was investigated in blood in both sexes from D5 to D29 as well as in the spleen, thymus and crural lymph nodes from females euthanized on D5 and D26. On D27, the cytokine response of male piglets to an injection of lipopolysaccharide was investigated. Special care was taken to minimize the stress at blood sampling and euthanasia. We found that early gestational stress only affected the relative weight of adrenals on D5 (S