Mild heat stress-induced adaptive immune response in blood mononuclear cells and leukocytes from mesenteric lymph nodes of primiparous lactating Holstein cows.

Heat stress negatively affects the metabolism and physiology of the bovine gut. However, it is not known whether heat stress induces an inflammatory response in mesenteric lymph nodes (MLN), the primary origin of gut immune cells, and thus contributes to inflammatory processes in the circulation. Therefore, our objective was to elucidate the effects of chronic heat stress on the systemic activation of acute-phase response in blood, proinflammatory cytokine production in peripheral blood mononuclear cells (PBMC), and the activation of the toll-like receptor signaling (TLR) 2/4 pathway in MLN leucocytes and their chemokines and chemokine receptor profiles in Holstein cows. Primiparous Holstein cows (n = 30; 169 ± 9 d in milk) were exposed to a temperature-humidity index (THI) of 60 [16°C, 63% relative humidity (RH)] for 6 d. Thereafter, cows were evenly assigned to 3 groups: heat-stressed (HS; 28°C, 50% RH, THI = 76), control (CON; 16°C, 69% RH, THI = 60), or pair-feeding (PF; 16°C, 69% RH, THI = 60) for 7 d. On d 6, PBMC were isolated and on d 7 MLN. Plasma haptoglobin, TNFα, and IFNγ concentrations increased more in HS than CON cows. Concomitantly, TNFA mRNA abundance was higher in PBMC and MLN leucocytes of HS than PF cows, whereas IFNG mRNA abundance tended to be higher in MLN leucocytes of HS than PF cows, but not for chemokines (CCL20, CCL25) or chemokine receptors (ITGB7, CCR6, CCR7, CCR9). Furthermore, the TLR2 protein expression tended to be more abundant in MLN leucocytes of HS than PF cows. These results suggest that heat stress induced an adaptive immune response in blood, PBMC, and MLN leukocytes involving the acute-phase protein haptoglobin, proinflammatory cytokine production, and TLR2 signaling in MLN leucocytes. However, chemokines regulating the leucocyte trafficking between MLN and gut seem not to be involved in the adaptive immune response to heat stress.

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.

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.

Intravenous lipid infusion affects dry matter intake, methane yield, and rumen bacteria structure in late-lactating Holstein cows.

Increasing the dietary fat content of ruminant diets decreases methane (CH4) production. This effect is caused by the toxic properties of fatty acids on rumen microbial populations, coating of feed particles diminishing the accessibility for microbes, and a reduction in dry matter intake (DMI). The latter effect is caused by postabsorptive long-chain fatty acids eliciting anorexic signaling; however, whether circulating long-chain fatty acids affect rumen CH4 production alike is unknown. To approach this question, 5 rumen-cannulated Holstein cows in late lactation received 2 jugular catheters and were kept in respiration chambers to measure CH4 production and DMI for 48 h. In a crossover design, cows were intravenously infused with a 20% lipid emulsion (LIPO) or 0.9% NaCl (CON). The LIPO cows received 2.1 kg of triglycerides/d [0.152 ± 0.007 g of triglycerides/(kg of BW × h)-1] consisting of 12.1% palmitic acid, 4.2% stearic acid, 31.1% oleic acid, and 52.7% linoleic acid. Blood and rumen fluid samples were taken hourly during the day. Results showed that LIPO compared with CON infusion increased plasma triglyceride as well as free fatty acid and serotonin concentrations but reduced the proportion of de novo synthesized milk fatty acids (sum of C6 to C16). Daily CH4 production and DMI were lower, whereas daily CH4 yield (CH4/DMI) was greater in LIPO than CON cows, although CH4 yield decreased from d 1 to d 2 by 2 to 14% in LIPO-infused cows only. This effect was associated with a higher (acetate + butyrate)/propionate ratio, tending lower propionate concentrations between 24 and 34 h of infusion, reduced relative abundances of genera belonging to Succinivibrio, Ruminococcaceae, and Ruminiclostridium, and greater relative Bacteroidetes genus abundances in the rumen.

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.

The Effects of Oral Quercetin Supplementation on Splanchnic Glucose Metabolism in 1-Week-Old Calves Depend on Diet after Birth.

BACKGROUND: Inadequate colostrum supply results in insufficient intake of macronutrients and bioactive factors, thereby impairing gastrointestinal development and the maturation of glucose metabolism in neonatal calves. The flavonoid quercetin has been shown to have health-promoting properties, including effects in diabetic animals. However, quercetin interacts with intestinal glucose absorption and might therefore exert negative effects in neonates. OBJECTIVE: We evaluated the interaction between neonatal diet and quercetin feeding on splanchnic glucose metabolism in neonatal calves. METHODS: Calves (n = 28) were assigned to 4 groups and fed either colostrum or a milk-based formula on days 1 and 2 and supplemented daily with 148 µmol quercetin aglycone/kg body weight [colostrum with quercetin (CQ+)/formula with quercetin (FQ+)] or without this substance [colostrum without quercetin (CQ-)/formula with quercetin (FQ-)] from days 2-8. From day 3 onward, all calves received milk replacer. A xylose absorption test was performed on day 3, and on day 7, blood samples were collected to study glucose first-pass uptake after [(13)C6]-glucose feeding and intravenous [6,6-(2)H2]-glucose bolus injection. Plasma concentrations of metabolites and hormones were measured by taking additional blood samples. A biopsy specimen of the liver was harvested on day 8 to measure the mRNA expression of gluconeogenic enzymes. RESULTS: Higher postprandial plasma concentrations of glucose, lactate, urea, adrenaline, noradrenaline, insulin, and glucagon on day 7 in colostrum-fed calves indicate that metabolic processes were stimulated. Postabsorptive xylose and glucose plasma concentrations each increased by an additional 26%, and splanchnic glucose turnover decreased by 35% in colostrum-fed calves, suggesting improved glucose absorption and lower splanchnic glucose utilization in colostrum-fed calves. Quercetin supplementation resulted in higher noradrenaline concentrations and enhanced peak absorption and oxidation of [(13)C6]-glucose by 10%. Liver mitochondrial phosphoenolpyruvate carboxykinase mRNA abundance was reduced by 34% in colostrum-deprived calves. CONCLUSIONS: Feeding colostrum during the first 2 d of life is crucial for maturation of splanchnic glucose metabolism in calves. Supplementing quercetin improves gastrointestinal absorption capacity, particularly in colostrum-deprived calves.

Effects of a 6-wk intraduodenal supplementation with quercetin on energy metabolism and indicators of liver damage in periparturient dairy cows.

Periparturient dairy cows experience metabolic challenges that result in a negative energy balance (EB) and a range of postpartum health problems. To compensate for the negative EB, cows mobilize fatty acids from adipose tissues, which can lead to fatty liver disease, a periparturient metabolic disorder. Flavonoids, such as quercetin (Q), are polyphenolic substances found in all higher plants and have hepatoprotective potential and the ability to prevent or reduce lipid accumulation in the liver. In ruminants, few studies on the metabolic effects of Q are available, and thus this study was conducted to determine whether Q has beneficial effects on EB, lipid metabolism, and hepatoprotective effects in periparturient dairy cows. Quercetin was supplemented intraduodenally to circumvent Q degradation in the rumen. Cows (n=10) with duodenal fistulas were monitored for 7wk. Beginning 3wk before expected calving, 5 cows were treated with 100mg of quercetin dihydrate per kilogram of body weight daily in a 0.9% sodium chloride solution for a total period of 6wk, whereas the control cows received only the sodium chloride solution. The plasma flavonoid levels were higher in the Q-treated cows than in the control cows. A tendency for higher postpartum (pp) than antepartum (ap) plasma flavonoid levels was observed in the Q-treated cows than in the controls, which was potentially caused by a reduced capacity to metabolize Q. However, the metabolic status of the Q-treated cows did not differ from that of the control cows. The pp increases in plasma aspartate aminotransferase and glutamate dehydrogenase activities were less in the Q-treated cows than in the control cows. The Q had no effect on energy expenditures, but from ap to pp the cows had a slight decline in respiratory quotients. Irrespective of the treatment group, the oxidation of fat peaked after calving, suggesting that the increase occurred because of an increased supply of fatty acids from lipomobilization. In conclusion, supplementation with Q resulted in lower pp plasma aminotransferase and glutamate dehydrogenase, which indicated reduced liver damage. However, the direct effects of Q on the liver and the implications for animal performance remain to be investigated.

Low and high dietary protein:carbohydrate ratios during pregnancy affect materno-fetal glucose metabolism in pigs.

Inadequate dietary protein during pregnancy causes intrauterine growth retardation. Whether this is related to altered maternal and fetal glucose metabolism was examined in pregnant sows comparing a high-protein:low-carbohydrate diet (HP-LC; 30% protein, 39% carbohydrates) with a moderately low-protein:high-carbohydrate diet (LP-HC; 6.5% protein, 68% carbohydrates) and the isoenergetic standard diet (ST; 12.1% protein, 60% carbohydrates). During late pregnancy, maternal and umbilical glucose metabolism and fetal hepatic mRNA expression of gluconeogenic enzymes were examined. During an i.v. glucose tolerance test (IVGTT), the LP-HC-fed sows had lower insulin concentrations and area under the curve (AUC), and higher glucose:insulin ratios than the ST- and the HP-LC-fed sows (P < 0.05). Insulin sensitivity and glucose clearance were higher in the LP-HC sows compared with ST sows (P < 0.05). Glucagon concentrations during postabsorptive conditions and IVGTT, and glucose AUC during IVGTT, were higher in the HP-LC group compared with the other groups (P < 0.001). (13)C glucose oxidation was lower in the HP-LC sows than in the ST and LP-HC sows (P < 0.05). The HP-LC fetuses were lighter and had a higher brain:liver ratio than the ST group (P < 0.05). The umbilical arterial inositol concentration was greater in the HP-LC group (P < 0.05) and overall small fetuses (230-572 g) had higher values than medium and heavy fetuses (≥573 g) (P < 0.05). Placental lactate release was lower in the LP-HC group than in the ST group (P < 0.05). Fetal glucose extraction tended to be lower in the LP-HC group than in the ST group (P = 0.07). In the HP-LC and LP-HC fetuses, hepatic mRNA expression of cytosolic phosphoenolpyruvate carboxykinase (PCK1) and glucose-6-phosphatase (G6PC) was higher than in the ST fetuses (P < 0.05). In conclusion, the HP-LC and LP-HC sows adapted by reducing glucose turnover and oxidation and having higher glucose utilization, respectively. The HP-LC and LP-HC fetuses adapted via prematurely expressed hepatic gluconeogenic enzymes.

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.

High-protein-low-carbohydrate diet during pregnancy alters maternal plasma amino acid concentration and placental amino acid extraction but not fetal plasma amino acids in pigs.

A high protein-low-carbohydrate diet during pregnancy can cause intra-uterine growth restriction. However, its impact during pregnancy on maternal, umbilical and fetal plasma amino acid (AA) profiles is unknown. A maternal high-protein (30 %)-low-carbohydrate (HP-LC) diet was compared with isoenergetic standard (12·1 % crude protein; ST) and low-protein (6·5 %)-high-carbohydrate (LP-HC) diets fed to nulliparous pregnant sows to examine changes in AA concentrations in maternal, venous and arterial umbilical and fetal plasma in mid and late pregnancy. At 64 and 94 days of pregnancy (dp), sows underwent Caesarean section, and maternal, umbilical and fetal plasma samples were collected. The HP-LC diet mainly affected maternal plasma AA concentrations. Plasma concentrations of Ile and Val were increased and those of Ala, Glu and Gly were decreased (P ≤ 0·05) in HP-LC compared with ST sows at 64 and 94 dp. The LP-HC diet decreased fetal plasma Glu concentration compared with the ST diet at 94 dp. Substantial AA catabolism was reflected by increased (P ≤ 0·05) maternal and fetal plasma urea concentrations with the HP-LC compared with the ST and LP-HC diets at 94 dp. Fractional placental extraction of Val was higher whereas those of Ala, Gln and Glu were lower in the HP-LC compared with the ST sows at 64 and 94 dp (P ≤ 0·05). Reduced fetal mass at 94 dp was accompanied by reduced fetal extraction of Lys and Pro in the HP-LC group (P ≤ 0·05). In conclusion, a maternal HP-LC diet during pregnancy altered maternal plasma composition of many AA and modified placental AA extraction to compensate for imbalanced maternal nutrient intake.

Limited and excess protein intake of pregnant gilts differently affects body composition and cellularity of skeletal muscle and subcutaneous adipose tissue of newborn and weanling piglets.

AIM: This study investigated whether dietary protein intake less (50%) or greater (250%) than requirements throughout gestation differently affects offspring body composition and cellular properties of skeletal muscle and subcutaneous adipose tissue (SCAT). METHODS: Primiparous gilts were fed iso-energetic diets containing adequate (22 AP), high (21 HP), or low (19 LP) protein contents. Newborn (n = 166) and weanling piglets cross-fostered to sows fed a standard diet (day 28; n = 83) were examined by morphological, biochemical, histological, and molecular analyses of the body, SCAT, and semitendinosus, longissimus, biceps femoris muscles. RESULTS: Lowered birth weight (BW) in response to the HP and LP diets (p < 0.01) resulted from decreases in all body constituents in LP, and mainly from reduced body fat in HP piglets (p < 0.05). In the light BW class within litters, HP piglets exhibited a greater percentage of muscle tissue (p < 0.05) than LP piglets. Less SCAT mass in HP and LP piglets resulted from reduced (p < 0.05) number, but not the size of adipocytes. The LP diet adversely affected myogenesis and muscular differentiation derived from less (p < 0.01) primary and secondary myofibers, lower creatine kinase activity (p < 0.05), less IGF2 mRNA (p < 0.10), and greater expression of the embryonic myosin heavy chain isoform (p < 0.01). Catch-up growth of LP but not HP pigs until day 28 increased body fat (p = 0.01). Despite compensated muscle growth in LP piglets, the deficit in myofiber number remained. CONCLUSION: Poor intrauterine environment by limited and excess protein supply retards fetal growth, but only limited protein supply impairs myogenesis, persistently restricts muscle growth potential, and favors obesity at infancy.

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.

Salivary Cortisol, but Not Oxytocin, Varies With Social Challenges in Domestic Pigs: Implications for Measuring Emotions.

Animals respond to inherently rewarding or punishing stimuli with changes in core affective states, which can be investigated with the aid of appropriate biomarkers. In this study we evaluate salivary cortisol (sCORT) and salivary oxytocin (sOXT) concentrations under baseline conditions and in response to two negatively- and two positively-valenced social challenges in 75 young pigs (Sus scrofa domesticus), housed and tested in eight social groups. We predicted that: (1) Relative to baseline, weaning and brief social isolation would be associated with increases in sCORT, due to psychosocial stress, and reductions in sOXT, due to a lack of opportunities for social support; and (2) Opportunities for social play, and reunions with group members after a separation would be associated with weaker sCORT responses, and increases in sOXT concentrations compared to baseline and to negative social challenges. Testing and sample collection occurred between 28 and 65 days of age and involved a within-subject design, in which every subject was sampled multiple times in neutral (baseline), negative and positive social contexts. We also recorded behavioral data and measured rates of agonism, play and affiliative interactions in the different contexts, prior to saliva sampling. As expected, negative social challenges were associated with robust cortisol responses. Relative to baseline, pigs also had higher sCORT responses to positive social challenges, although these differences were only significant during reunions. Salivary oxytocin concentrations did not differ between the different social conditions, although sOXT was lowest during the brief social isolation. Behavioral analyses confirmed predictions about the expected changes in social interactions in different social contexts, with increases in agonism following weaning, increases in coordinated locomotor play in the play context and high rates of affiliative interactions during reunions. Relative sCORT reactivity to different contexts may reflect the intensity of emotional responses, with greater increases occurring in response to challenges that involve more psychosocial stress. Our results suggest that sOXT is not a reliable indicator of emotional valence in pigs, although more research is needed to characterize sOXT responses to various challenges with and without access to social support.

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.

Tryptophan metabolism, from nutrition to potential therapeutic applications.

Tryptophan is an indispensable amino acid that should to be supplied by dietary protein. Apart from its incorporation into body proteins, tryptophan is the precursor for serotonin, an important neuromediator, and for kynurenine, an intermediary metabolite of a complex metabolic pathway ending with niacin, CO(2), and kynurenic and xanthurenic acids. Tryptophan metabolism within different tissues is associated with numerous physiological functions. The liver regulates tryptophan homeostasis through degrading tryptophan in excess. Tryptophan degradation into kynurenine by immune cells plays a crucial role in the regulation of immune response during infections, inflammations and pregnancy. Serotonin is synthesized from tryptophan in the gut and also in the brain, where tryptophan availability is known to influence the sensitivity to mood disorders. In the present review, we discuss the major functions of tryptophan and its role in the regulation of growth, mood, behavior and immune responses with regard to the low availability of this amino acid and the competition between tissues and metabolic pathways for tryptophan utilization.

The Effect of Dietary Protein Imbalance during Pregnancy on the Growth, Metabolism and Circulatory Metabolome of Neonatal and Weaned Juvenile Porcine Offspring.

Protein imbalance during pregnancy affects women in underdeveloped and developing countries and is associated with compromised offspring growth and an increased risk of metabolic diseases in later life. We studied in a porcine model the glucose and urea metabolism, and circulatory hormone and metabolite profile of offspring exposed during gestation, to maternal isoenergetic low-high (LP-HC), high-low (HP-LC) or adequate (AP) protein-carbohydrate ratio diets. At birth, LP-HC were lighter and the plasma acetylcarnitine to free carnitine ratios at 1 day of life was lower compared to AP offspring. Plasma urea concentrations were lower in 1 day old LP-HC offspring than HP-LC. In the juvenile period, increased insulin concentrations were observed in LP-HC and HP-LC offspring compared to AP, as was body weight from HP-LC compared to LP-HC. Plasma triglyceride concentrations were lower in 80 than 1 day old HP-LC offspring, and glucagon concentrations lower in 80 than 1 day old AP and HP-LC offspring. Plasma urea and the ratio of glucagon to insulin were lower in all 80 than 1 day old offspring. Aminoacyl-tRNA, arginine and phenylalanine, tyrosine and tryptophan metabolism, histidine and beta-alanine metabolism differed between 1 and 80 day old AP and HP-LC offspring. Maternal protein imbalance throughout pregnancy did not result in significant consequences in offspring metabolism compared to AP, indicating enormous plasticity by the placenta and developing offspring.

Time-Dependent Effects of Acute Handling on the Brain Monoamine System of the Salmonid Coregonus maraena.

The immediate stress response involves the activation of the monoaminergic neurotransmitter systems including serotonin, dopamine and noradrenaline in particular areas of the fish brain. We chose maraena whitefish as a stress-sensitive salmonid species to investigate the influence of acute and chronic handling on the neurochemistry of monoamines in the brain. Plasma cortisol was quantified to assess the activation of the stress axis. In addition, we analyzed the expression of 37 genes related to the monoamine system to identify genes that could be used as markers of neurophysiological stress effects. Brain neurochemistry responded to a single handling (1 min netting and chasing) with increased serotonergic activity 3 h post-challenge. This was accompanied by a modulated expression of monoaminergic receptor genes in the hindbrain and a significant increase of plasma cortisol. The initial response was compensated by an increased monoamine synthesis at 24 h post-challenge, combined with the modulated expression of serotonin-receptor genes and plasma cortisol concentrations returning to control levels. After 10 days of repeated handling (1 min per day), we detected a slightly increased noradrenaline synthesis and a down-regulated expression of dopamine-receptor genes without effect on plasma cortisol levels. In conclusion, the changes in serotonergic neurochemistry and selected gene-expression profiles, together with the initial plasma cortisol variation, indicate an acute response and a subsequent recovery phase with signs of habituation after 10 days of daily exposure to handling. Based on the basal expression patterns of particular genes and their significant regulation upon handling conditions, we suggest a group of genes as potential biomarkers that indicate handling stress on the brain monoamine systems.