Effects of 2 liquid feeding rates over the first 3 months of life on whole-body energy metabolism and energy use efficiency of dairy calves up to 5 months.

Intensified milk replacer (MR) feeding in calves has nutritional long-term effects and is suggested to increase milk production later in life. However, the underlying mechanisms are not completely understood. The aim of our study was to investigate whether MR feeding intensity has long-term effects on energy metabolism and energy use efficiency of dairy calves. Newborn female Holstein calves (n = 28) were randomly assigned to 2 liquid feeding groups offered daily either 10% of body weight (BW) colostrum followed by 10% of BW MR (10%-MR) or 12% of BW colostrum followed by 20% of BW MR (20%-MR). Calves were housed individually. Weaning was completed by the end of wk 12. Hay and calf starter were fed from d 1 until the end of wk 14 and 16, respectively. A total mixed ration was fed from wk 11 onward, and the metabolizable energy intake (MEI) was determined daily. Energy metabolism of calves was measured in respiratory chambers before weaning in wk 6 and 9, and after weaning in wk 14 and 22. The MEI/BW0.75 was higher before weaning but lower during and shortly after weaning in 20%-MR calves. During the preweaning period, the 20%-MR animals had higher average daily gain, BW, back fat thickness and muscle diameter, but lower plasma ß-hydroxybutyrate concentrations. The group difference in average daily gain ceased in wk 9, differences in back fat thickness and muscle diameter ceased after weaning, whereas difference in BW0.75 persisted until wk 23. The energy conversion ratio (BW gain/MEI) was not different before weaning, but was lower during and after weaning in 20%-MR calves. The higher MEI and BW0.75 in 20%-MR calves resulted in higher heat production (HP), as well as in higher carbohydrate oxidation (COX) and fat oxidation during the preweaning period. Gas exchange variables normalized to BW0.75 or MEI differed between groups only during preweaning. The energy balance was lower in 10%-MR calves in wk 6 and 9. The HP/BW0.75 and COX/BW0.75 were higher, whereas HP/MEI was lower in 20%-MR calves in wk 6. When normalized to BW0.75 and MEI, HP in wk 6 and 9, and COX in wk 9 was lower in 20%-MR calves. In conclusion, 20%-MR calves showed greater efficiency estimates preweaning, but this effect did not occur after weaning, suggesting that energy use efficiency does not persist until later stages in life.

A natural Ala610Val substitution causing glucocorticoid receptor hypersensitivity aggravates consequences of endotoxemia.

Despite the crucial role of glucocorticoid receptor (GR) in proper immune responses, the effect of GR hypersensitivity on inflammation is rarely reported. To fill this knowledge gap, we exploited the natural gain-of-function substitution in the porcine glucocorticoid receptor (GRAla610Val) and challenged pigs carrying normal or hypersensitive GR using 50 µg/kg lipopolysaccharide (LPS) following pretreatment with either saline or single bolus of 60 µg/kg dexamethasone (DEX). The GRAla610Val substitution reduced baseline cortisol, adrenocorticotropic hormone (ACTH), and triglyceride concentration and granulocyte proportion whereas baseline platelet counts were elevated. Val-carriers, i.e. AlaVal as well as ValVal pigs, showed less LPS-induced cortisol rise but the cortisol fold change was similar in all genotypes. Differently, ACTH response to LPS was most significant in GRAla610Val heterozygotes (AlaVal). LPS-induced disorders, including sickness behaviors, anorexia, thrombocytopenia, cytokine production, and metabolic alterations were more intense in Val-carriers. On the other hand, Val-carriers were more sensitive to DEX effect than wild types (AlaAla) during endotoxemia, but not under unchallenged conditions. This is the first report revealing aggravated responses to endotoxemia by GR gain-of-function. Together, these results imply that GR hypersensitivity is difficult to diagnose but may represent a risk factor for endotoxemia and sepsis.

Is it getting in the hair? - Cortisol concentrations in native, regrown and segmented hairs of cattle and pigs after repeated ACTH administrations.

Adrenocorticotropic hormone (ACTH) is part of the hypothalamic-pituitary-adrenal axis response to stress and induces the release of cortisol, which is commonly used as an indicator in stress and animal welfare research. In recent years, hair cortisol concentration (HCC) gained increasing importance as a promising retrospective indicator for stress in animals. Thus, the aim of our study was to validate HCC as a potential indicator of increased endogenous cortisol release in cattle and pigs by repeated ACTH administrations followed by cortisol analysis in different hair types. For this purpose, 34 cattle and 38 gilts were treated either with repeated i.m. injections of ACTH or saline every second day over a period of 4 weeks. Saliva samples were taken before and after injections once a week from selected animals to verify the endogenous cortisol response. At the end of the treatment (week 4) and after 8 and 12 weeks, samples of natural and regrown hair were taken from the caudo-dorsal region of the back and analyzed for cortisol concentrations. In addition, natural hair was sampled after 12 weeks and cut into segments prior to analysis. Treatment with ACTH revealed a significant increase in salivary cortisol after application in both species, although this increase was attenuated in pigs compared to cattle. In week 4, HCCs were significantly elevated in natural and regrown hair of ACTH-treated animals. In cattle, HCCs significantly increased after ACTH treatment in natural, regrown and segmental hair compared with control animals, indicating that HCC may be a promising indicator of stress, as cortisol levels in all hair types reflected the preceding period with increased cortisol release. In pigs, there were no differences in HCCs between treatments. This may be caused by the lower systemic cortisol response in pigs, but seasonally reduced hair growth and external cross-contamination of hair by saliva and urine under commercial husbandry conditions may also interfere with the validity of HCC in this species.

It's getting hairy - External contamination may affect the validity of hair cortisol as an indicator of stress in pigs and cattle.

Hair cortisol concentration (HCC) is increasingly used for the assessment of enhanced hypothalamic-pituitary-adrenal (HPA) axis activity, e.g., caused by repeated or long-term stressful conditions or pathologies. However, there is still a lack of knowledge regarding the mechanisms and sources of cortisol incorporation into the hair and possible confounding factors, especially in non-human animals. Farm animals are usually kept under confined housing conditions, have close contact with each other and with soiled environments and may thus be exposed to contamination with urine, feces and saliva, which are known to contain substantial concentrations of cortisol or its metabolites. Therefore, the aim of our study was to investigate the impact of contamination with urine, feces and saliva on the cortisol concentration in the hair of pigs and cattle. In an in vitro experiment, hair strands of 12 pigs and 12 cattle were repeatedly contaminated with urine and saliva, containing either low or high cortisol concentrations, or with the feces of the respective species and were compared with hair treated with water or untreated hair. Contamination was performed over 20 days for two hours daily. Thereafter, all samples were washed, ground, extracted and analyzed for HCCs following the same protocol. Our results showed that contamination with urine caused a considerable and concentration-dependent increase in HCCs in both species. Saliva had a comparable effect only in cattle. In addition, the treatment with water led to a reduction in the cortisol concentration of porcine hair, whereas contamination with feces caused an increase in HCC only in cattle. Our findings provide evidence that contamination of hair with cortisol-containing body fluids causes incorporation of cortisol into the hair shaft, probably via diffusion depending on the concentration gradient. In that case, cortisol in hair derived from contamination cannot be distinguished from cortisol originating from blood. Thus, contamination may affect the validity of hair cortisol as an indicator of HPA axis activity and cannot be prevented by decontamination protocols prior to analysis.

Within a hair's breadth - Factors influencing hair cortisol levels in pigs and cattle.

During the last two decades, hair cortisol concentration (HCC) has proven to be a promising marker for the evaluation of increased hypothalamic-pituitary-adrenal axis activity caused by repeated or long-term stressful conditions. A minimally invasive sampling procedure, simple storage and the retrospective characteristic of one hair sample are reasons why HCC is increasingly used not only in human medicine but also in animal welfare research. However, before applying HCC as a reliable indicator for stress, it is important to investigate potential influencing factors in addition to stressors in the species of interest. Thus, the aim of our study was to elucidate the impact of age, sex, hair color, body region, age of hair segments and season of hair sampling on HCC in pigs and cattle. Hair samples were taken by electric clippers and analyzed by ELISA after extraction. Our results show similar effects of influencing factors in both species. Significantly increased HCCs were found in young animals after birth compared with older age groups. In addition, HCCs were significantly higher in samples obtained from the tail tip in comparison with samples from the shoulder, neck and back regions, in black hair compared with white hair and in distal hair segments. Season had an impact on HCC only in cattle, which exhibited higher levels in winter than in summer. In conclusion, age, body region, hair color, hair segment and season affect hair cortisol concentrations and should be considered and controlled for when HCC is applied as a potential stress indicator in pigs and cattle. In addition, further research is necessary to elucidate the mechanisms by which cortisol is incorporated into the hair shaft.

Early-Life Maternal Deprivation Predicts Stronger Sickness Behaviour and Reduced Immune Responses to Acute Endotoxaemia in a Pig Model.

Early-life adversity may have programming effects on neuroendocrine and immune adaptation mechanisms in humans and socially living animals. Using a pig model, we investigated the effect of daily 2-h maternal and littermate deprivation from postnatal days 2-15, either alone (DA) or in a group of littermates (DG) on the neuroendocrine, immunological and behavioural responses of piglets challenged with the bacterial endotoxin lipopolysaccharide (LPS) on day 42. LPS increased plasma concentrations of cortisol, tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) and induced typical signs of sickness in all piglets. DA+DG piglets showed stronger signs of sickness compared to control (C) piglets. Plasma TNF-α concentrations were significantly lower in DA+DG males. In addition, the TNF-α/IL-10 ratio was significantly lower in DA than in DG and C males. Gene expression analyses showed lower hypothalamic TNF-α mRNA expression and diminished mRNA expression of the mineralocorticoid receptor (MR) and IL-10 in the amygdala of DA+DG piglets in response to LPS. Interestingly, males showed a higher MR- and a lower IL-10 mRNA expression in the amygdala than females. The present data suggest that repeated maternal deprivation during early life may alter neuroendocrine and immune responses to acute endotoxaemia in a sex-specific manner.

The use of hair cortisol for the assessment of stress in animals.

The hair cortisol concentration (HCC) is assumed to be a retrospective marker of integrated cortisol secretion and stress over longer periods of time. Its quantification is increasingly used in psychoneuroendocrinological studies in humans, but also in animal stress and welfare research. The measurement of HCCs for the assessment of stress offers many considerable benefits for use in domesticated and wild animals, especially due to the easy and minimally invasive sampling procedure and the representation of longer time periods in one sample. This review aims to outline the different fields of application and to assess the applicability and validity of HCC as an indicator for chronic stress or long-term activity of the hypothalamic-pituitary-adrenal axis in wild and domesticated animals. Specific hair characteristics are presented and the advantages and limitations of using HCC are discussed. An overview of findings on the impact of stress- and health-related factors on HCCs and of diverse influencing factors causing variation in hair cortisol levels in different species is given. Recommendations for the use of hair cortisol analysis are proposed and potential fields of future research are pointed out. The studies indicate an effect of age and pregnancy on HCCs, and cortisol incorporation into hair was also found to depend on hair colour, body region, sex and season of year, but these results are less consistent. Furthermore, the results in animals show that a wide array of stressors and pathological conditions alters the cortisol concentrations in hair and that HCC thereby provides a reliable and valid reflection of long-term cortisol secretion in many species. However, more research is necessary to investigate the underlying mechanisms of cortisol incorporation into the hair and to explore the hair growth characteristics in the species of interest. To overcome confounding influences, the use of standardized sampling protocols is strongly advised.

Kinetics of Physiological and Behavioural Responses in Endotoxemic Pigs with or without Dexamethasone Treatment.

Although dexamethasone (DEX) is a widely used immunoregulatory agent, knowledge about its pharmacological properties in farm animals, especially pigs, is insufficient. Previous studies suggest that compared to other species, pigs are less sensitive to the immunosuppression conferred by DEX and more sensitive to the threat of bacterial endotoxins. However, there is a paucity of studies examining DEX immunomodulation in endotoxemia in this species. In this study, a porcine endotoxemia model was established by lipopolysaccharide (LPS) and the effect of DEX-pretreatment on the magnitude and kinetics of neuroendocrine, metabolic, hematologic, inflammatory, and behavioural responses were examined. DEX decreased cortisol, adrenocorticotropic hormone (ACTH), red blood cell, hemoglobin, hematocrit, and lymphocyte whereas glucose concentration was increased under both normal and endotoxemic conditions. By contrast, DEX decreased triglyceride, lactate, and IL-6 concentrations and increased platelet count only under an endotoxemic condition. DEX also reduced the frequency of sickness behaviour following LPS challenge. PCA showed that glucose and triglyceride metabolism together with red blood cell count mainly contributed to the separation of clusters during DEX treatment. Our study demonstrates that DEX protects pigs from inflammation and morbidity in endotoxemia, in spite of their less sensitivity to DEX. Moreover, its considerable role in the regulation of the metabolic and hematologic responses in endotoxemic pigs is revealed for the first time.

Psychosocial stress sensitizes neuroendocrine and inflammatory responses to Escherichia coli challenge in domestic piglets.

Exposure to psychosocial stress can have a profound impact on immune reactivity and health mediated by hypothalamic-pituitaryadrenal (HPA) axis activation. However, current knowledge regarding the mechanisms involved in cross-sensitization between stress and the immune system is limited. Here, we investigated the effects of a single social isolation followed by repeated oral Escherichia coli (E. coli) applications on cortisol, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), haptoglobin and C-reactive protein (CRP) concentrations in the blood; on clinical signs of disease; and on mRNA expression of the glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11ß-hydroxysteroid dehydrogenase 1 and 2 (11ß-HSD1 and 11ß-HSD2), TNF-α and IL-6 in the hypothalamus, prefrontal cortex (PFC) and spleen of 7-, 21- and 35-day-old piglets. Additionally, the protein levels of splenic TNF-α and IL-6 were analyzed. Non-isolated, E. coli-challenged piglets served as a control. Social isolation for 4 h induced a rise in the plasma cortisol concentrations immediately after social treatment and after repeated E. coli applications in isolated compared to non-isolated piglets. The circulating TNF-α concentration was not affected by social treatment. Furthermore, previously isolated piglets showed a higher frequency of signs of disease in response to E. coli challenge than non-isolated piglets, while the haptoglobin and CRP concentrations did not significantly differ between social treatments. In the brain, 11ß-HSD1, 11ß-HSD2 and IL-6 mRNA expression in the hypothalamus and GR, and 11ß-HSD1 and 11ß-HSD2 mRNA expression in the PFC were higher in isolated, E. coli-challenged piglets than in the corresponding controls. Moreover, isolated piglets also displayed higher MR, 11ß-HSD1 and IL-6 mRNA expression levels and TNF-α concentrations in the spleen. The stress-induced alterations in the hypothalamus and spleen were particularly pronounced in younger piglets. The present findings may contribute to a better understanding of the complex interplay between early psychological stress and an increased risk of disease and might also have implications on aspects of the health and welfare of farm animals and humans.

Sea Buckthorn Pomace Supplementation in the Diet of Growing Pigs-Effects on Fatty Acid Metabolism, HPA Activity and Immune Status.

There is evidence that sea buckthorn, as a source of n-3 polyunsaturated fatty acids (n-3 PUFA), possesses health-enhancing properties and may modulate neuroendocrine and immune functions. In the present study, we investigated the effect of sea buckthorn pomace (SBP) supplementation in the diet of growing German Landrace pigs on fatty acids in the blood and hypothalamus, peripheral immune parameters and mRNA expression of corticotropin-releasing hormone (CRH), mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the hypothalamus and spleen. Pigs were fed diets supplemented with 12% of dried SBP or 0% SBP (control group) over an intervention period of eight weeks. The fatty acid profiles in blood plasma were significantly affected by SBP supplementation only for C18:2n-6 and n-6/n-3 PUFA ratio compared with the control group. SBP supplementation did not significantly affect the fatty acid concentrations in the hypothalamus. Furthermore, there were no significant differences in mRNA expression of CRH, MR and GR in the hypothalamus or of GR mRNA expression in the spleen. Concerning the immune status, the plasma IgG levels tended to be higher in SBP pigs, whereas the leukocyte distribution, mitogen-stimulated lymphocyte proliferation, and serum IgM levels remained unchanged. In conclusion, the SBP supplementation of the diet only caused moderate effects on fatty acid metabolism, but no significant effects on hypothalamic-pituitary-adrenal (HPA) activity and immunity in growing pigs. It seems that a beneficial effect of dietary n-3 PUFA on health and welfare is more likely to be expected during stressful situations.

Effects of 1-Methyltryptophan on Immune Responses and the Kynurenine Pathway after Lipopolysaccharide Challenge in Pigs.

An enhanced indoleamine 2,3-dioxygenase 1 (IDO1) activity is associated with an increased mortality risk in sepsis patients. Thus, the preventive inhibition of IDO1 activity may be a promising strategy to attenuate the severity of septic shock. 1-methyltryptophan (1-MT) is currently in the interest of research due to its potential inhibitory effects on IDO1 and immunomodulatory properties. The present study aims to investigate the protective and immunomodulatory effects of 1-methyltryptophan against endotoxin-induced shock in a porcine in vivo model. Effects of 1-MT were determined on lipopolysaccharide (LPS)-induced tryptophan (TRP) degradation, immune response and sickness behaviour. 1-MT increased TRP and its metabolite kynurenic acid (KYNA) in plasma and tissues, suppressed the LPS-induced maturation of neutrophils and increased inactivity of the animals. 1-MT did not inhibit the LPS-induced degradation of TRP to kynurenine (KYN)-a marker for IDO1 activity-although the increase in KYNA indicates that degradation to one branch of the KYN pathway is facilitated. In conclusion, our findings provide no evidence for IDO1 inhibition but reveal the side effects of 1-MT that may result from the proven interference of KYNA and 1-MT with aryl hydrocarbon receptor signalling. These effects should be considered for therapeutic applications of 1-MT.

Effects of social support on glucocorticoid sensitivity of lymphocytes in socially deprived piglets.

There is growing evidence that social support given by a conspecific attenuates stress responses of a socially deprived animal. We hypothesized that the presence of a familiar social partner modulates the effectiveness of social buffering as assessed by an altered glucocorticoid sensitivity of immune cells. The current study investigated the effects of a 4-h social deprivation procedure on stress hormone responses and immune cell functions in 7-, 21- and 35-day-old piglets (52 males and 56 females). Within each of the three age categories, nine piglets were deprived of their mother and littermates either alone or with a familiar or unfamiliar age-matched piglet. Compared to non-deprived controls, piglets that were alone displayed significant increases in plasma adrenocorticotropic hormone and cortisol concentrations, and all socially deprived piglets showed a greater in vitro proliferative response of peripheral blood mononuclear cells (PBMCs) to lipopolysaccharide (LPS)-stimulation than controls. Concanavalin A (ConA)-induced in vitro proliferation was not affected by social treatment. Additionally, both the ConA- and LPS-stimulated PBMCs from piglets that experienced any of the deprivation treatments were more resistant to the inhibitory effects of cortisol than PBMCs from the controls in each of the three age categories. Irrespective of the mitogen used, the presence of an age-matched conspecific during deprivation attenuated the dose-dependent cortisol resistance. Here, familiarity between the piglets clearly improved the effectiveness of social support in an age-dependent manner. Collectively, our findings emphasize the benefits of social partners regarding stress coping abilities, with positive implications for animal welfare and health.

Social support attenuates the adverse consequences of social deprivation stress in domestic piglets.

Social deprivation is a severe stressor affecting a number of behavioral and physiological functions of gregarious species. It is assumed that, dependent upon the level of familiarity, social support given by a conspecific may attenuate the adverse consequences of stress. We investigated the effects of a 4h maternal and littermate deprivation on behavioral reactions, stress hormone responses and brain corticosteroid receptor expression in 7-, 21- and 35-day-old domestic piglets (Sus scrofa) that were left alone or in the presence either of a familiar or unfamiliar age-matched piglet. Compared to control animals, all of the socially deprived piglets showed significant stress responses, such as impaired habituation in repeated open-field/novel-object tests, enhanced ACTH and cortisol release, and altered corticosteroid receptor expression in the hypothalamus. In addition, our results demonstrated that younger piglets had more difficulty coping with stress. The presence of an age-matched conspecific had a direct calming effect on the deprived piglet during the deprivation procedure, which was revealed by diminished stress-induced HPA activity and altered reactions in the behavioral test situations (e.g., activity, escape, and vocalization). Furthermore, because the presence of a familiar piglet causes a more pronounced buffering effect, we have shown for the first time that the degree of familiarity between the piglets may influence the effectiveness of social support. Our study emphasizes the benefits of social partners on positive welfare and the ability for pigs to cope with stress; therefore, our results should be taken into account during handling practices such as weaning and mixing.

Hair cortisol concentration in postpartum dairy cows and its association with parameters of milk production.

Hair cortisol concentration (HCC) is considered as an indicator for a minimally invasive assessment of long-term stress. In dairy cows, in addition to stress influences, changing physiological conditions during gestation and lactation (eg, due to varying energy requirements or fluctuating milk yield) may affect HCCs. Thus, the aim of our study was to investigate HCCs of dairy cows during different stages of lactation and to determine the relationship between milk production traits and hair cortisol levels. Samples of natural hair and regrown hair were collected from 41 multiparous Holstein Friesian cows at 100-d intervals from parturition to 300 d postpartum. All samples were analyzed for cortisol concentration and the association of HCC with milk productions traits was evaluated. Our results show that cortisol concentration in natural hair increased after parturition and was highest 200 d postpartum. Cumulative milk yield from parturition to 300 d showed moderate and positive correlation with HCC in natural hair at 300 d. There was a positive correlation between urea concentration in milk and cortisol levels in regrown hair at 200 d, and between somatic cell count in milk and HCC in natural and regrown hairs 200 d postpartum. Together, these findings suggest that physiological loads during lactation, eg, caused by metabolic stress and/or inflammation, may be associated with increased HCC levels. In addition, the results on hair color confirm previous findings in cattle that black hair has higher cortisol concentrations than white hair. Black hair therefore appears to be more suitable for hair cortisol analysis as it provides higher protection against photodegradation.

Tumour necrosis factor receptor deficiency alters anxiety-like behavioural and neuroendocrine stress responses of mice.

Tumour necrosis factor (TNF)-alpha is known to be involved in anxiety and the regulation of the hypothalamic-pituitary-adrenal axis. To examine the role of its receptors in neuroendocrine immunomodulation, we studied behaviour, corticosterone production and T-cell activation in mice with a C57BL/6J background and deficient for one or both TNF receptors (TNFR1-/-, TNFR2-/-, and TNFR1+2-/-) compared to wildtype C57BL/6J mice with and without psychological stress. Stress was induced by social disruption (SDR), and anxiety-like behaviour was examined using the elevated plus maze (EPM). Anxiety of unstressed TNFR1+2-/- mice was increased compared to C57BL/6J mice as shown by reduced ratios of entries into open arms relatively to total entries. SDR-stressed TNFR1+2-/- mice showed reduced ratios of entries into open arms relatively to total entries, reduced ratios of distances walked in open relatively to distances walked in both arms and reduced time in open arms compared to C57BL/6J mice. Locomotor activity of unstressed and SDR-stressed TNFR1-/- and TNFR2-/- mice was reduced. Serum corticosterone concentrations of control mice do not differ between mouse strains. However, TNFR1+2-/- mice had significantly higher corticosterone concentrations than C57BL/6J mice after SDR. EPM testing significantly increased corticosterone concentrations in all strains. Mitogen-induced activation-marker expression was reduced in TNFR1-/- T-helper cells under control and stress conditions, while activation marker expression of TNFR2-/- and TNFR1+2-/- cells was only slightly affected by stress compared to C57BL/6J T cells. Our study suggests that both TNF receptors contribute to anxiety-like behaviour and corticosterone responses, whereas TNFR1 has a larger impact on T-cell activation.

Effects of inadequate maternal dietary protein:carbohydrate ratios during pregnancy on offspring immunity in pigs.

BACKGROUND: Inadequate nutrition in utero may retard foetal growth and alter physiological development of offspring. This study investigated the effects of low and high protein diets fed to primiparous German Landrace sows throughout pregnancy on the immune function of their offspring at different ages. Sows were fed diets with adequate (AP, 12.1%; n = 13), low (LP, 6.5%; n = 15), or high (HP, 30%; n = 14) protein content, made isoenergetic by varying carbohydrate levels. Cortisol, total protein and immunoglobulin (IgG, IgM, IgA) concentrations were measured in the blood of sows over the course of pregnancy. Cortisol, total protein, immunoglobulins, lymphocyte proliferation, immune cell counts, and cytokines were assessed in the blood of offspring at baseline and under challenging conditions (weaning; lipopolysaccharide (LPS) administration). RESULTS: In sows, the LP diet increased cortisol (P < 0.05) and decreased protein levels (P < 0.01) at the end of pregnancy. Immunoglobulin concentrations were decreased in LP (IgA) and HP piglets (IgG, IgM and IgA) on the first day of life (P < 0.05), whereas the number of lymphocytes and mitogen-induced lymphocyte proliferation of the piglets were unaffected by the maternal diet. Mortality during the suckling period was higher in LP piglets compared with AP and HP offspring (P < 0.01). Furthermore, LP piglets showed an elevated cortisol response to weaning, and in HP piglets, the CD4+ cell percentage and the CD4+/CD8+ ratio increased after weaning (P < 0.05). The lipopolysaccharide-induced rise of IL-6 was higher in LP (P = 0.09) and HP (P < 0.01) compared with AP piglets, and LP piglets displayed higher IL-10 levels than AP piglets (P < 0.05). CONCLUSIONS: Our results indicate that both low and high protein:carbohydrate ratios in the diet of pregnant sows can induce short-term as well as long-lasting effects on immune competence in piglets that may have serious consequences for host defence against bacterial pathogens.

Early-life maternal deprivation affects the mother-offspring relationship in domestic pigs, as well as the neuroendocrine development and coping behavior of piglets.

Early-life adversity may have programming effects on the psychological and physiological development of offspring. Domestic pigs (Sus scrofa) are an excellent model species for studying these effects because of their many physiological similarities to humans. Piglets from 10 sows were subjected to daily 2-h maternal deprivation on postnatal days (PND) 2-15 alone (DA) or in a group of littermates (DG). Control piglets (C) from 10 sows stayed with their mothers. Mother-offspring interaction, milk oxytocin, and cortisol were analyzed. An open-field/novel-object (OF/NO) test was performed with piglets on PNDs 16 and 40. Plasma cortisol and immune parameters were determined on PND 5 and 16. Two piglets from each group and sow were sacrificed on PND 20 and stress-related gene expression in the limbic system and prefrontal cortex (PFC), as well as splenic lymphocyte proliferative abilities, were examined. The milk cortisol of sows increased during the first separation of mother and offspring on the second day of lactation, whereas milk oxytocin did not change. The increase in cortisol by the OF/NO test on PND 16 was greater in C piglets than in DA and DG ones. DA piglets showed less agitated behavior than DG and C piglets in the OF/NO test at PND 16, but appeared more fearful. On PND 40, DA piglets showed more arousal than DG and C piglets in the OF/NO test. Neither plasma IgA nor N/L ratios in blood nor mitogen-induced proliferation of spleen lymphocytes were affected by deprivation. We found a higher mRNA expression of CRHR1 in the hypothalamus and a higher expression of MR in the hippocampus in DA piglets than in DG ones. The expression of GR, MR, and CRHR1 genes in the PFC was reduced by maternal deprivation, however, the expression of arginine vasopressin and oxytocin receptors was not affected. Repeated maternal deprivation induces sustained effects on stress reactivity and behavior of domestic piglets. Some of these effects were buffered by the presence of littermates. In addition, we found sex-specific differences in behavior and gene expression.

Age-related changes in corticosteroid receptor expression and monoamine neurotransmitter concentrations in various brain regions of postnatal pigs.

Negative early life experience may be associated with altered functioning of stress-related systems and may increase vulnerability to diseases later in life. Corticosteroids are important mediators of homeostasis and stress and exert their effects via two receptors, the mineralocorticoid receptor (MR) and the glucocorticoid receptor (GR), and through the glucocorticoid-metabolizing enzymes 11ß-hydroxysteroid dehydrogenase (11ß-HSD) types 1 and 2 in a brain-region-specific manner. However, relatively little is known about the postnatal ontogeny of these receptors and enzymes in the central nervous system. Here we describe, for the first time, the postnatal ontogeny of central GR, MR, 11ß-HSD1, and 11ß-HSD2 gene expression and monoamine levels in stress-related brain regions of domestic pigs at 7, 21, and 35 days of age. During the postnatal period, there was an increase in GR, MR, and 11ß-HSD1 mRNA expression in the pituitary and prefrontal cortex and an increase in MR mRNA expression in the hippocampus. We also demonstrated age-dependent changes in levels of noradrenaline and dopamine and their metabolites in the locus coeruleus, with the highest concentrations on day 7 compared with days 21 and 35. In conclusion, the dynamic changes in corticosteroid receptors and monoamines during neural development of postnatal pigs may represent periods of sensitivity to environmental stress that are comparable to some extent with those that are observed in primates and humans. Thus, these findings support the use of the domestic pig as an alternative animal model for humans in stress research.

Intestinal glucose absorption but not endogenous glucose production differs between colostrum- and formula-fed neonatal calves.

Glucose supply markedly changes during the transition to extrauterine life. In this study, we investigated diet effects on glucose metabolism in neonatal calves. Calves were fed colostrum (C; n = 7) or milk-based formula (F; n = 7) with similar nutrient content up to d 4 of life. Blood plasma samples were taken daily before feeding and 2 h after feeding on d 4 to measure glucose, lactate, nonesterified fatty acids, protein, urea, insulin, glucagon, and cortisol concentrations. On d 2, additional blood samples were taken to measure glucose first-pass uptake (FPU) and turnover by oral [U-(13)C]-glucose and i.v. [6,6-(2)H(2)]-glucose infusion. On d 3, endogenous glucose production and gluconeogenesis were determined by i.v. [U-(13)C]-glucose and oral deuterated water administration after overnight feed deprivation. Liver tissue was obtained 2 h after feeding on d 4 and glycogen concentration and activities and mRNA abundance of gluconeogenic enzymes were measured. Plasma glucose and protein concentrations and hepatic glycogen concentration were higher (P < 0.05), whereas plasma urea, glucagon, and cortisol (d 2) concentrations as well as hepatic pyruvate carboxylase mRNA level and activity were lower (P < 0.05) in group C than in group F. Orally administered [U-(13)C]-glucose in blood was higher (P < 0.05) but FPU tended to be lower (P < 0.1) in group C than in group F. The improved glucose status in group C resulted from enhanced oral glucose absorption. Metabolic and endocrine changes pointed to elevated amino acid degradation in group F, presumably to provide substrates to meet energy requirements and to compensate for impaired oral glucose uptake.

Altered immunomodulation by glucocorticoids in neonatal pigs exposed to a psychosocial stressor.

Stressful early life experiences can have short- and long-term effects on neuroendocrine and immune mechanisms of adaptation, which are primarily modulated by glucocorticoids. This study aimed to examine how the stress and immune systems interact to cope with psychosocial stress induced by a single social isolation (4 h) in neonatal pigs at 7, 21, or 35 d of age. This social isolation provoked increased plasma ACTH and cortisol concentrations and reduced TNF-α levels but had no significant effect on IL-6 levels. Socially isolated piglets had a higher lipopolysaccharide (LPS)-stimulated proliferative response of peripheral blood mononuclear cells (PBMCs) than controls, whereas concanavalin A (ConA)-induced proliferation was not affected by isolation. A single social isolation also induced a dose-dependent cortisol resistance in ConA- and LPS-stimulated PBMCs compared with controls, which may be an adaptive response in the short term. Moreover, LPS-stimulated cultures from control piglets showed a reduction in cortisol sensitivity with increasing age. Conclusively, these findings provide stress-related measures for the psychophysiological assessment of livestock handling practices but might also have implications for stress and health studies in young animals and humans.