The effects of thermal environment and spray-dried plasma on the acute-phase response of pigs challenged with lipopolysaccharide

Forty barrows (TR4 x C22) were weaned at 17 d of age (BW = 6.27 +/- 0.30 kg), housed (two pigs/pen) in a thermal-neutral environment (TN; constant 26.7 degrees C), and fed diets with or without 7% (as-fed basis) spray-dried plasma (SDP). On d 7, one pig/ pen was moved into a cold environment (CE; constant 15.6 degrees C). Pigs were fitted with jugular catheters on d 11. On d 12, 16 pigs per environment (eight pigs per dietary treatment) were challenged i.v. with 75 microg of lipopolysaccharide (LPS)/kg of BW. Blood samples were collected over a 4.5-h period. Pigs were then killed and tissue samples were harvested for messenger RNA (mRNA) analysis. From d 0 to 7, pigs fed SDP diets had a lower gain:feed ratio (G/F) than pigs fed no SDP (533 +/- 14 vs. 585 +/- 17 g/kg; P < 0.03). Pigs housed in the CE consumed more feed and had a lower G/F than pigs housed in TN from d 7 to 11 (P < 0.001). There were no environment x diet interactions from d 7 to 11 (P > 0.78). Baseline concentrations of serum ACTH and cortisol were lower in the TN pigs than in the CE pigs (P < 0.001). Pigs fed diets without SDP had lower serum cortisol concentrations over the 4.5-h period than pigs fed SDP (time x diet, P < 0.001). Serum concentrations of tumor necrosis factor-alpha (TNF-alpha) were highest for pigs consuming SDP in the CE, whereas there were no differences among the other treatments (time x diet x environment, P < 0.02). Pigs housed in the CE had higher serum interleukin-1beta (IL-1beta) (P < 0.001) and interleukin-6 (IL-6; P < 0.001) than TN pigs. Pigs fed SDP also had slightly higher serum IL-1beta concentrations (P < 0.10) and higher (P < 0.001) IL-6 concentrations than pigs fed no SDP. Pigs fed SDP had 9% lower liver and 13% lower thymus mRNA expression of tumor necrosis factor-alpha (TNF-alpha) than pigs that consumed no SDP (P < 0.06). Liver IL-1beta, IL-6, and LPS-binding protein mRNA were higher in the CE than in the TN (P < 0.03, P < 0.001, and P < 0.05; respectively). In addition, spleen TNF-alpha (P < 0.03) and IL-6 (P < 0.01) mRNA levels were higher in the CE than in the TN. Pigs consuming SDP and challenged with LPS responded with elevated serum concentrations of cortisol and cytokines compared with pigs fed diets with no SDP. Housing pigs in a CE increased the baseline concentrations of ACTH and cortisol, and when coupled with an LPS challenge, resulted in elevated serum and tissue mRNA levels of cytokines. Cold stress and feeding SDP during a LPS challenge may result in increased stress and immune responses in young pigs.     

Evaluation of immune system function in neonatal pigs born vaginally or by Cesarean section

Full term crossbred sows were selected to study the interaction of the immune system, hypothalamus-pituitary-adrenal axis, and growth in pigs born by Cesarean section (c-section; n=4 sows) or vaginal birth (n=4 sows). Gestation length and birth weight did not differ between vaginal birth and c-section pigs (P=0.34 and 0.62, respectively). Blood and tissue samples were collected from 44 pigs at birth. Forty-five pigs were weaned at 13 d. On d 14, pigs received an i.p. injection of lipopolysaccaride (LPS; 150 microg/kg) or saline at min 0, and blood samples were collected at -20, -10, 0, 5, 10, 20, 40, 60, 90, and 120 min. Vaginal birth pigs had 21% greater average daily gain than c-section pigs on d 14 (P<0.01). Basal serum concentrations of adrenocorticotrophin (ACTH) and cortisol were greater in c-section than vaginal birth pigs at birth (P<0.01) but were not different at 14 d (P=0.99 and 0.80, respectively). LPS increased serum concentrations of ACTH, cortisol, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha; P<0.01) but the response was not different between c-section and vaginal birth (P>0.22). Basal serum concentrations of TNF-alpha tended to be greater in c-section vs vaginal birth pigs at 14 d (P=0.0967); however, basal serum concentrations of IFN-gamma tended to be lower in c-section pigs vs vaginal birth pigs at 14 d (P=0.0787). Expression of interleukin (IL)-6, IL-6 receptor, IL-1beta, and TNF-alpha mRNA did not differ between vaginal birth and c-section pigs but changed in an age and tissue dependent manner. Thus, reduced growth rate of c-section pigs is associated with altered immune system function.     

Weaned pig responses to Escherichia coli K88 oral challenge when receiving a lysozyme supplement

Lysozyme is a low-molecular-weight protein with antimicrobial properties. An experiment was conducted to investigate the response of piglets receiving a water-soluble lysozyme supplement [Entegard (EG), Neova Technologies Inc., Abbotsford, British Columbia, Canada; 4,000 lysozyme units/mg] after oral challenge with enterotoxigenic Escherichia coli (ETEC). A total of 36 individually housed weanling pigs were randomly allotted to 1 of the 4 treatments, with 9 replicates per treatment. Treatments were a control (CONT, no additive), antibiotic (AB; 2.5 g/kg of feed of antibiotic with chlortetracycline, sulfamethazine, and penicillin), and EG delivered in the drinking water at concentrations of 0.1% (EG1) and 0.2% (EG2). All pigs received a basal diet similar in composition and nutrients, except for pigs receiving the AB diet, which had an added antibiotic. Pigs were acclimated to treatments for a 7-d period to monitor growth performance. On d 8, blood samples were collected from each pig to obtain serum, and each pig was gavaged with 6 mL (2 × 10(9) cfu/mL) of ETEC solution. Pigs were monitored for another 7 d to assess incidences of diarrhea and growth performance, and then all pigs were killed to obtain intestinal tissue and digesta samples. Treatments did not influence growth performance throughout the study. Greater ETEC counts were observed in the ileal mucosal scrapings (P = 0.001) and colonic digesta (P = 0.025) of pigs in the CONT group compared with pigs in the AB and EG1 groups. Pigs receiving AB and EG1 had greater (P < 0.05) small intestinal weights and ileal villus heights than pigs receiving CONT; however, the ileal villus height-to-crypt depth ratio was greater in pigs fed the AB diet (1.69) compared with those fed the CONT diet (1.34), whereas pigs receiving EG1 were intermediate. Pigs in the EG1 group showed greater (P < 0.001) serum tumor necrosis factor α and IL-6 concentrations before ETEC challenge; however, at 7 d postchallenge, pigs receiving EG2 showed the least (P < 0.05) circulating tumor necrosis factor α and IL-6 concentrations. Overall, better intestinal growth and development, as well as decreased ETEC counts on the intestinal mucosa and serum proinflammatory cytokines, suggest that EG can maintain gut health and function in piglets commensurate with antibiotics. However, it is noteworthy that at the largest dose tested, EG seemed to have a dramatic effect on proinflammatory cytokines but had a minimal or no effect on the other response criteria.     

Effect of sodium butyrate on growth performance and response to lipopolysaccharide in weanling pigs

Two experiments were conducted to determine the effects of dietary sodium butyrate on growth performance and response to Escherichia coli lipopolysaccharide (LPS) in weanling pigs. In a 28-d experiment, 180 pigs (initial BW 6.3 kg) were fed 0, 0.05, 0.1, 0.2, or 0.4% sodium butyrate, or 110 mg/kg of dietary tylosin. There was no effect of dietary sodium butyrate or tylosin on overall G:F, but there was a linear trend (P < 0.07) toward decreased ADFI and ADG as levels of sodium butyrate increased. In a second 28-d experiment, 108 pigs (initial BW 6.3 kg) were assigned to 1 of 3 dietary treatments: 1) no antibiotics, 2) 0.2% sodium butyrate, or 3) 55 mg/kg of carbadox. On d 14, a subset of pigs from the no-antibiotic and butyrate treatment groups was challenged with E. coli LPS or injected with sterile saline in a 2 x 2 factorial arrangement (+/-LPS challenge; +/-dietary butyrate; n = 6 pigs/treatment group). Four hours after LPS challenge, blood samples were obtained, and samples of LM, liver, and ileum were collected for gene expression analysis. Serum samples were analyzed for IL-6, tumor necrosis factor alpha (TNFalpha), alpha(1)-acid glycoprotein, cortisol, IGF-I, insulin, and metabolites. The relative abundance of tissue cytokine and IGF-I mRNA was measured by real-time PCR. Feeding diets containing sodium butyrate or carbadox did not alter ADG or ADFI compared with pigs fed the control diet. From d 0 to 14, pigs fed diets containing 0.2% sodium butyrate had decreased (P < 0.05) ADG and tended (P < 0.06) to have decreased G:F compared with animals fed diets containing carbadox. Challenge with LPS increased (P < 0.05) serum cytokines and cortisol and decreased (P < 0.05) serum glucose and triglycerides. Injection with LPS increased (P < 0.05) the relative abundance of hepatic IL-6 and TNFalpha mRNA, increased (P < 0.05) LM TNFalpha mRNA content, and decreased (P < 0.05) IGF-I mRNA in LM. For serum cortisol, there was an interaction (P < 0.05) between dietary butyrate and LPS. The increase in serum cortisol attributable to LPS was greater (P < 0.05) in pigs fed butyrate than in pigs fed the control diet. There tended (P < 0.10) to be an interaction between LPS and diet and for butyrate to increase the relative abundance of IL-6 mRNA in LM. Carbadox did not alter cytokine or IGF-I mRNA or serum metabolites, but did decrease (P < 0.05) serum TNFalpha. These data indicate that dietary sodium butyrate does not enhance growth performance, but may regulate the response to inflammatory stimuli in weanling pigs.     

Duration of feeding linseed diet influences expression of inflammation-related genes and growth performance of growing-finishing barrows

The aim of the study was to investigate the effects of dietary linseed (rich in n-3 PUFA) on expression of inflammation-related genes and on growth performance of growing-finishing barrows. Two isoenergetic and isonitrogenous diets were formulated, one as the basal diet and the other containing 10% linseed. Twenty-four Landrace x Yorkshire barrows weighing 35 +/- 3.7 kg were randomly assigned to 1 of 4 treatment groups, with 6 pigs per group. During the entire experimental period of 90 d, these 4 groups of pigs were first fed the basal diet and then fed the linseed diet for 0, 30, 60, and 90 d before slaughter, respectively. Pig growth; messenger RNA (mRNA) expression of peroxisome proliferator-activated receptor-gamma (PPARgamma), IL-1beta0, IL-6, and tumor necrosis factor-alpha (TNF-alpha); and plasma concentrations of the 3 proinflammatory cytokines were measured and analyzed. Average daily feed intake did not differ among treatment groups (P > 0.05), but ADG (P < 0.05) and G:F (P < 0.01) responded quadratically to the duration of linseed diet feeding, and pigs in the 60-d treatment group had the greatest ADG and G:F. The mRNA expression of PPARgamma in loin muscle and spleen increased linearly (P < 0.01) with the duration of linseed diet feeding, whereas its expression in adipose tissue was not affected (P = 0.095). Tumor necrosis factor-alpha and IL-6 mRNA expression in muscle, adipose, and spleen, as well as serum concentration of TNF-alpha, decreased linearly (P < 0.01) with the duration of linseed diet feeding. Peroxisome proliferator-activated receptor-gamma mRNA abundance was negatively correlated with IL-1beta, IL-6, and TNF-alpha mRNA abundance both in muscle (R(2) = 0.63, P < 0.001) and in spleen (R(2) = 0.69, P < 0.001), and PPARgamma mRNA expression in spleen (R(2) = 0.59, P < 0.01) and muscle (R(2) = 0.52, P < 0.05) was negatively correlated with serum TNF-alpha concentration. There were also significant quadratic relations between ADG and expression of PPARgamma (P < 0.05) and splenic TNF-alpha (P < 0.05). These data suggest that intake of n-3 PUFA from the linseed diet led to significant decreases in the expression of proinflammatory cytokine genes, which may stimulate growth in growing-finishing barrows, at least in part, through a PPARgamma-dependent mechanism.     

Hormonal profiles, behavioral responses, and short-term growth performance after castration of pigs at three, six, nine, or twelve days of age

The objective of this study was to determine the effects of castration on short-term growth performance, hormone profiles, and behavior in pigs at 3, 6, 9, or 12 d of age. Ninety intact male pigs were assigned randomly to a treatment age by litter [3, 6, 9, or 12 d of age; n = 9 to 13 pigs per treatment (age) group]. Pigs within a single litter were then assigned to noncastrated (NC) or castrated (CAS) treatment groups according to BW. Pigs were nonsurgically fitted with jugular catheters, and blood samples were drawn immediately before castration (0 h) and at 0.5, 1, 1.5, 2, 24, and 48 h after castration. Body weights were obtained when pigs were catheterized and again at 24 and 48 h after castration. Serum samples were analyzed for cortisol, porcine corticosteroid-binding globulin, and dehydroepiandrosterone sulfate (DHEA-S). No differences were detected in initial BW of pigs, and there was no overall treatment effect on growth performance of pigs at 24 or 48 h posttreatment. A time x treatment interaction was detected (P < 0.01) for serum cortisol concentrations, such that cortisol was greater in CAS pigs than in NC pigs. No overall effect of age at castration was observed on cortisol concentrations. At 24 h after castration, serum cortisol concentrations returned to baseline in all treatment groups; however, at 48 h after castration, overall cortisol concentrations were elevated (P < 0.01) in the 6-, 9-, and 12-d-old pigs in both the CAS and NC groups compared with baseline concentrations. Total cortisol and porcine corticosteroid-binding globulin were used to calculate the free cortisol index (FCI). A time x treatment interaction was observed (P < 0.01) for FCI, such that FCI was greater in CAS males than in NC males. The FCI was also affected by age (P < 0.01). There was a time x treatment x age interaction (P < 0.01) for serum DHEA-S, such that DHEA-S concentrations decreased in CAS animals but increased in NC animals, and DHEA-S concentrations increased with age. During the first 2 h after castration, there was an overall age effect (P = 0.01) on the time that pigs spent standing, such that 3-d-old pigs stood more than 6-, 9-, or 12-d-old pigs. Treatment did not influence the time that pigs spent nursing, lying, standing, or sitting, although there was a trend (P = 0.08) for CAS pigs to be less active than NC pigs. These data indicate that castration is stressful regardless of age; however, the stress associated with handling seems to increase as pigs age.     

Effects of feeding diets containing bacitracin methylene disalicylate to heat-stressed finishing pigs

The objective of this study was to evaluate the effects of heat stress and dietary bacitracin methylene disalicylate (BMD) on growth performance, carcass characteristics, and immunological responses in finishing pigs. Four groups of 32 finishing pigs (n = 128) with initial BW between 80 to 90 kg were used. Pigs were fed a corn-soybean meal-distillers grains-based control or BMD (31.5 mg/kg) diet for a 14-d adaptation period at the thermal neutral temperature (23°C), and continued to be fed their respective diets when exposed to a constant temperature (23°C) or a cyclical heat stress environment (37°C from 1000 to 1900 h and 27°C from 1900 to 1000 h) for a 28-d experimental period. Each group of pigs was housed in 4 rooms, with 2 pens/room and 4 pigs/pen. Saliva samples from each pig were collected on d -1 (initial baseline), 1, 13, and 27 for cortisol analysis. Concentrations of haptoglobin, IL-1β, and tumor necrosis factor-α were determined in serum samples on d -1, 1, 13, and 27. Pigs exposed to heat stress had 31% less ADG (P < 0.001), 23% less ADFI (P < 0.001), 9% less G:F (P < 0.001), and 34% greater average daily water intake (P = 0.03) than those in the non-heat-stress conditions. Dietary BMD tended to reduce ADG (P < 0.07) compared with the control (0.66 vs. 0.73 kg/d, respectively). Heat stress increased (P < 0.05) saliva cortisol on d 1, but no effects were observed on subsequent days. Serum haptoglobin concentrations were greater (P < 0.05) in heat-stressed pigs on d 1, and concentrations tended to remain greater (P < 0.1) on d 13. Pigs fed the BMD diet tended to have a longer villus height (P = 0.07) in the duodenum and greater crypt depths in the duodenum (P = 0.09) and jejunum (P = 0.07). Heat-stressed pigs tended to have a decreased proportion of propionate (P = 0.08), greater acetate:propionate (P = 0.08), and a reduced proportion of valerate (P = 0.02) in the cecum. These results indicate that BMD did not counteract the negative effects of heat stress on growth performance, but BMD appears to increase villus height and crypt depth in the duodenum. Furthermore, heat stress appears to alter VFA production in finishing pigs.     

Effects of beta-glucan obtained from the Chinese herb Astragalus membranaceus and lipopolysaccharide challenge on performance, immunological, adrenal, and somatotropic responses of weanling pigs

A total of 108 crossbred piglets (7.75 +/- 0.24 kg of BW) weaned at 28 d was used to study the interactive effects of beta-glucan obtained from the Chinese herb Astragalus membranaceus (AM) and Escherichia coli lipopolysaccharide (LPS) challenge on performance, immunological, adrenal, and somatotropic responses of weaned pigs. The treatments were in a 2 x 3 factorial arrangement; main effects were level of Astragalus membranaceus glucan (AMG; 0, 500, or 1,000 mg/kg; as-fed basis) and presence of immunological challenge (with or without LPS). The experiment included six replicate pens per treatment and three pigs per pen. Lipopolysaccharide challenges were conducted on d 7 and 21 of the trial. Blood samples were obtained from the vena cava from one pig per pen at 3 h after LPS challenge to determine plasma responses. Weight gain and feed:gain ratio were unaffected by glucan. However, there was a quadratic effect on feed intake (P < 0.05): pigs fed 500 mg of glucan/kg had the highest feed intake. Immunological challenge with LPS decreased weight gain (P = 0.02). An interaction (P = 0.01 to 0.09) between AMG and LPS was observed for glucose, IL-1beta, PGE2, and cortisol. Astragalus membranaceus glucan had a quadratic effect on the plasma concentrations of glucose, IL-1beta, PGE2, and cortisol (P < 0.05) after both LPS challenges. Plasma concentrations of glucose, IL-1beta, PGE2, and cortisol (P < 0.05) were all increased in LPS-challenged pigs compared with the control pigs after both LPS challenges. The IGF-I concentrations were less for LPS-challenged pigs than for unchallenged pigs. The lymphocyte proliferation response of peripheral blood induced by 5 microg of concanavalin A/mL (P < 0.01) and IL-2 bioactivity (P < 0.05) increased linearly with increasing addition of glucan. Pigs challenged with LPS had greater T-lymphocyte proliferation (P = 0.06) and IL-2 bioactivity (P = 0.07) than unchallenged pigs after the first immunological challenge but not after the second. In conclusion, although glucan did not improve pig performance under the conditions of the present experiment, when included at 500 mg/kg, it decreased the release of inflammatory cytokine and corticosteroid and improved the lymphocyte proliferation response of weanling piglets via enhanced IL-2 bioactivity.     

The influence of intramammary lipopolysaccharide infusion on serum Ca, P, vitamin D, cytokines and cortisol concentrations in lactating sows

Ten multiparous lactating sows were used to investigate whether intramammary infusion of lipopolysaccharides (LPS; Escherichia coli 0111:B4; 2.0 microg/kg of body weight) would affect the circulating concentrations of Ca, P, 25-hydroxyvitamin D (25-OHD), tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and cortisol. The sows were randomly allotted to either control group (control) or LPS-treated group with five individuals per group and were infused with either physiological saline solution or LPS solution. The rectal temperature and udder quarter appearance were recorded at 0 (just before infusion), 1, 3, 7, 12 or 24 h after infusion. Blood samples were taken at 0, 1, 3, 7, 12 or 24 h after infusion. Before infusion, the rectal temperatures of all sows were below 39.2 degrees C. At 3 and 7 h after infusion, the sows in the LPS group had a rectal temperature over 39.4 degrees C. At 24 h after infusion, the rectal temperatures returned to pre-infusion levels. Serum Ca and P concentrations in the LPS group decreased (P < 0.05) after LPS infusion compared with the control group at 1 h after infusion. No significant differences (P > 0.05) in the concentrations of 25-OHD were observed between groups control and LPS at any sampling time. Increased (P < 0.01) concentrations of serum TNF-alpha, IL-6 and cortisol were observed in the LPS group compared with the control group at 3 and 7 h after infusion respectively. In conclusion, the elevation of serum concentrations of TNF-alpha, IL-6 and cortisol and the alterations of circulating concentrations of Ca and P following LPS infusion indicate that the immune system has been activated and immune activation may affect macromineral homeostatic regulation, which might have important implications for metabolic health of lactating sows. Lowered serum Ca and P following immune activation also shows a causative mechanism whereby immune activation increases the risk of secondary disorders such as mastitis-metritis-agalactia syndrome. However, immune activation did not affect circulating concentrations of vitamin D metabolites.     

Effect of spray-dried plasma and lipopolysaccharide exposure on weaned pigs: II. Effects on the hypothalamic-pituitary-adrenal axis of weaned pigs

A study was conducted with 20 weaned barrows (14 d, 4.98 +/- 0.21 kg) to determine the effect of feeding spray-dried plasma (SDP) after weaning on the pig's stress response to a lipopolysaccharide (LPS) challenge. After weaning, pigs were fed a diet containing 0 or 7% SDP for 7 d. On d 6 after weaning, all pigs were nonsurgically fitted with a jugular catheter. On d 7 after weaning, the pigs were given i.p. injections of either saline or LPS (150 microg/kg BW) followed by serial blood collection every 15 min for a 3-h period. Following the 3-h blood collection, all pigs were killed and tissue was collected for mRNA analysis. Pig weight on d 7 after weaning was not affected by dietary treatment (P > 0.21). Pigs fed the diet with SDP had lower (P < 0.05) levels of hypothalamic corticotropin-releasing hormone (CRH) mRNA, pituitary gland CRH receptor mRNA, and adrenal gland adrenocorticotropin-releasing hormone (ACTH) receptor mRNA. Dietary treatment did not affect pituitary gland proopiomelanocortin (POMC) mRNA. No effect of LPS treatment was observed in any of the mRNA levels examined. For both serum ACTH and cortisol, there was a significant diet x LPS treatment interaction (P < 0.01) such that both the ACTH and cortisol responses to the LPS challenge were greater in the pigs fed the diet with SDP than in the pigs fed the diet without SDP. For pigs given the saline injection, diet did not affect basal serum cortisol concentration; however, basal serum ACTH concentration was lower in those pigs fed the diet with SDP (P < 0.0001). A diet x LPS treatment interaction (P < 0.024) was observed for adrenal gland mRNA expression for steroidogenic acute regulatory (StAR) protein such that the LPS-induced increase in StAR mRNA was greater in the pigs fed SDP than in pigs fed the diet without SDP. These results demonstrate that pigs fed a diet with SDP have an increased activation of the pituitary-adrenal axis following an LPS challenge compared to pigs fed a diet without SDP.     

Supplemental vitamin C and yeast cell wall beta-glucan as growth enhancers in newborn pigs and as immunomodulators after an endotoxin challenge after weaning

To test possible dietary immune modulators, 32 crossbred male pigs were given 1 of 4 dietary treatments (8 pigs/treatment): control, Saccharomyces cerevisiae with beta-glucan (Energy Plus, Natural Chem Industries LTD, Houston, TX; 0.312 g/kg of BW, 2.5% of diet), vitamin C (Stay C 35, DSM Nutritional Products Inc., Prisippany, NJ; 75 ppm), or beta-glucan plus vitamin C together (combination; 0.312 g/kg of BW and 75 ppm, respectively). Supplements were given in whole milk within 36 h of birth and then daily for 2 wk until weaning, when the supplement was given in feed for an additional 2 wk. Growth was recorded during the 4 wk of supplement delivery. An i.v. lipopolysaccharide challenge (LPS; 150 microg/kg) was given 14 d postweaning at 0900. Behavior was observed, and blood samples were collected every 30 min for 4 h via a jugular catheter from -1 (0800) to 3 (1200) h relative to challenge (-60, -30, 0, 30, 60, 90, 120, 150, and 180 min), and tissues were collected after exsanguination. Beta-glucan (glucan and combination) increased (P < 0.05) BW and ADG compared with vitamin C and control. Cortisol concentrations showed an interaction (P < 0.05) of the beta-glucan and vitamin C. Intestinal expression of tumor-necrosis factor (TNF)-alpha mRNA was greatest for vitamin C and beta-glucan compared with control and combination, and liver TNF-alpha mRNA expression showed a main effect (P < 0.01) of beta-glucan. Lung expression of TNF-alpha mRNA exhibited a vitamin C effect (P < 0.01). In contrast, spleen had greater (P < 0.01) relative abundance of TNF-alpha mRNA in beta-glucan pigs. Intestinal expression of IL-1Ra mRNA was greater (P < 0.05) for vitamin C and beta-glucan treatments compared with the control and combination pigs. Liver expression of IL-1 receptor antagonist mRNA exhibited a vitamin C effect (P < 0.01). Lying and sleeping behaviors differed (P < 0.05) among treatments early in the observations (0700 to 0720), then sporadically until 50 min after the LPS injection. The vitamin C group slept less (P < 0.05) on those occasions. The time spent lying was least (P < 0.05) for the glucan and combination pigs immediately after the injection. These results show a complex interaction between vitamin C and this yeast product after LPS challenge, with differential expression in tissues by 2 h after LPS injections. The combination enhanced postweaning growth and reduced TNF-alpha expression of the intestinal and liver tissues, suggesting an important immunomodulatory role of the combination treatment.     

In utero heat stress alters the postnatal innate immune response of pigs

The effects of in utero heat stress (IUHS) range from decreased growth performance to altered behavior, but the long-term impact of IUHS on postnatal innate immune function in pigs is unknown. Therefore, the study objective was to determine the effects of early gestation IUHS on the immune, metabolic, and stress response of pigs subjected to an 8 hr lipopolysaccharide (LPS) challenge during postnatal life. Twenty-four pregnant gilts were exposed to thermoneutral (TN; n = 12; 17.5 ± 2.1 °C) or heat stress (HS; n = 12; cyclic 26 to 36 °C) conditions from days 6 to 59 of gestation, and then TN conditions (20.9 ± 2.3 °C) from day 60 of gestation to farrowing. At 12 wk of age, 16 IUHS and 16 in utero thermoneutral (IUTN) pigs were selected, balanced by sex and given an intravenous injection of LPS (2 µg/kg BW mixed with sterile saline [SAL] and injected at 2 µL/kg BW) or SAL (2 µL/kg BW). Body temperature was monitored every 30 min, and blood was obtained at 0, 1, 2, 3, 4, 6, and 8 hr following the LPS challenge. Blood samples were analyzed for glucose, insulin, non-esterified fatty acids (NEFA), cortisol, and cytokine concentrations. In addition, white blood cell counts were determined at 0 and 4 hr. Hour 0 data were used as covariates. Body temperature was increased (P < 0.01) in LPS (40.88 ± 0.08 °C) vs. SAL (39.83 ± 0.08 °C) pigs. Eosinophils tended to be decreased overall (P = 0.09; 43.9%) in IUHS vs. IUTN pigs. Glucose concentrations were reduced overall (P = 0.05; 5.9%) in IUHS vs. IUTN pigs. The NEFA concentrations tended to be greater (P = 0.07; 143.4%) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs tended to have greater (127.4%) circulating NEFA concentrations compared with IUTN-SAL and IUHS-SAL pigs. Cortisol was increased (P = 0.04) in IUHS-LPS compared with IUTN-LPS pigs at 3 hr (21.5%) and 4 hr (64.3%). At 1 hr, tumor necrosis factor α was increased (P = 0.01; 115.1%) in IUHS-LPS compared with IUTN-LPS pigs. Overall, interleukin-1β (IL-1β) and interleukin-6 (IL-6) were greater (P < 0.04; 281.3% and 297.8%, respectively) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs had increased IL-1β and IL-6 concentrations compared with IUTN-SAL and IUHS-SAL pigs. In summary, IUHS altered the postnatal cytokine, metabolic, and physiological stress response of pigs during postnatal life, which may have negative implications toward the innate immune response of IUHS pigs to pathogens.     

Birth by caesarian section alters postnatal function of the hypothalamic-pituitary-adrenal axis in young pigs

Eight crossbred sows were selected for the present study (n = 4 vaginal delivery and n = 4 Caesarian section [C-section]). Gestation length did not differ between vaginal delivery and C-section pigs (113.6 +/- .1 and 113.2 +/- .3 d, respectively; P > .16). Blood and tissue samples from 38 pigs were collected at birth. All remaining pigs were sustained with vaginal-delivery sows until 2 wk of age (n = 39). At 2 wk of age, remaining pigs were catheterized for blood sample collection to assess pituitary-adrenal responsiveness to an injection of corticotropin-releasing hormone (CRH; 10 microg/kg). Blood samples were collected at -30, -15, 0, 5, 10, 20, 40, 60, and 90 min; pigs received CRH or saline at time 0. Pigs were killed and tissue samples were collected immediately following the last blood sample. Serum concentrations of ACTH and cortisol (CS) were measured. Total RNA was isolated from the pituitary and adrenal glands to evaluate gene expression for mRNA specific for pro-opiomelanocortin (POMC) and for the ACTH receptor. Centrifuged clot:blood ratio was reduced in the C-section pigs at birth (P < .001) and at 2 wk of age (P < .043), compared with the vaginally delivered pigs. Basal serum concentration of ACTH was greater in C-section than in vaginally delivered pigs at birth (P = .01) but did not differ at 2 wk of age (P = .42). Basal serum concentration of CS was not different at birth (P = .86) but was greater in C-section pigs than in vaginally delivered pigs at 2 wk of age (P < .04). Serum concentration of ACTH was not different (P > .99) between the two groups of pigs following the CRH challenge. However, serum concentration of CS was greater (P < .05) in C-section pigs following the CRH challenge. Expression of ACTH receptor mRNA tended to be greater in C-section pigs at birth (P < .063) and at 2 wk of age (P < .016). There was no difference in POMC mRNA between treatments (P > .73); however, there was a developmental increase (P < .001) from birth to 2 wk of age. These data indicate that the birth process plays an important role in postnatal function of the hypothalamic-pituitary-adrenal axis in young pigs.     

Cytokine mRNA profiles in pigs exposed prenatally and postnatally to Schistosoma japonicum

The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed pigs and 4 uninfected control pigs were compared 27 weeks post exposure. Variables included worm burdens, tissue egg counts, liver pathology and mRNA levels of IL-2, IL-4, IL-10, IL-12, TNF-alpha, TGF-beta1 and IFN-gamma in the liver and the caecum, assessed by RT-PCR. Infection intensity and level of septal fibrosis were significantly higher in the postnatal group compared to the prenatal group (P < 0.05). A significant increase of IL-4 (P < 0.01), IL-10 (P < 0.01), IL-12 (P < 0.01) and TNF-alpha (P < 0.05) mRNA level was also observed in the caecum of prenatally infected animals compared to the control group (P < 0.01). The prenatal group showed higher levels of TGF-beta1 in the liver compared with the postnatally infected group (P< 0.05) and the control group (P< 0.01). This suppressive immune response correlated with previously reported low hepatic pathogenesis in prenatally exposed pigs.     

Interleukin-6 and insulin incrase and nitric oxide and adiponectin decrease in blind dogs with pituitary-dependent hyperadrenocorticism

In this study, two populations of dogs with pituitary dependent hypercortisolism (PDH) were compared over a 2-year period. One group had normal vision (Group A, n=27) and one group was blind (Group B, n=20). Group B was characterised by the rapid appearance of the clinical signs of PDH that precede blindness. We found increases in pre-adrenocorticotropic hormone cortisol (P=0.002), IL-6 (P=0.0001), insulin, and insulin sensitivity (detected with the Homeostatic Model Assessment, P<0.0001) in Group B but not in Group A. The nitric oxide (NO) and the total adiponectin concentrations decreased (P=0.0001 and P=0.02, respectively) in Group B versus Group A. The IL-6 and insulin concentrations and the HOMA-A index were positively correlated with the cortisol concentration and were negatively correlated with the NO concentration. With the exception of adiponectin, the other variables were associated with blindness. We concluded that blindness in PDH is a haemodynamic event associated with metabolic changes, with the increase in the IL-6 concentration and the decrease in the NO concentration affecting the retinal vasculature and producing a high risk of vision loss.     

The effect of flaxseed supplementation on growth, carcass characteristics, fatty acid profile, retail shelf life, and sensory characteristics of beef from steers finished on grasslands of the northern Great Plains

The objective of this trial was to determine if daily supplementation of flaxseed for 85 d to steers finished on grasslands of the northern Great Plains would influence growth and carcass characteristics or the fatty acid profile, tenderness, and sensory characteristics of beef steaks. Eighteen Angus yearling steers (initial BW 399 ± 21 kg) were randomly divided into 3 groups. Steers in treatment 1 (FLX; n = 6) received a daily supplement of ground flaxseed (0.20% of BW), whereas steers in treatment 2 (CSBM; n = 6) received a daily supplement of ground corn and soybean meal (0.28% of BW), with contents of CP and TDN being similar to the supplement for FLX. Control steers (CONT; n = 6) were not supplemented. Treatments were given to each individual steer in side-by-side stalls and were fed from mid-August to November 7, 2007, the day before slaughter. All steers grazed growing forage from early May through the first week of November. Growth rate of steers fed FLX was 25% greater (P < 0.01) than that of steers fed CONT, but was similar (P = 0.45) to that of steers fed CSBM. No differences were observed for carcass characteristics (P ≥ 0.14), tenderness (Warner-Bratzler shear force; P ≥ 0.24), or sensory attributes (P ≥ 0.40) except for a slight off-flavor detected in steaks from steers fed FLX compared with CONT (7.4 vs. 7.8, respectively, with 8 indicating no off-flavor and 1 indicating extreme off-flavor; P = 0.07) and CSBM (7.9; P = 0.01). The n-3 fatty acids α-linolenic acid and eicosapentaenoic acid were 62 and 22% greater, respectively, in beef from steers fed FLX compared with those fed CONT (P < 0.001). The ratio of n-6 to n-3 fatty acids was smaller (P < 0.001) in beef from steers fed FLX compared with the ratios in beef from steers fed CONT and CSBM. Daily supplementation of flaxseed to steers grazing growing vegetation on the northern Great Plains may improve growth rate and enhance the n-3 fatty acid profile of the steaks.     

Effects of exogenous ghrelin on feed intake, weight gain, behavior, and endocrine responses in weanling pigs

The objectives were to determine relative ADG, ADFI, behavior, and endocrine responses in weaned pigs receiving exogenous ghrelin. Twenty-four barrows weaned at 18 d of age (d 0 of the experiment) were catheterized via the jugular vein, weighed, and assigned to either a ghrelin (n = 12) or saline (control; n = 12) infusion group. Initial pig BW did not differ between treatments (7.87+/-0.39 vs. 7.92+/-0.35 kg for ghrelin and control treatments, respectively). Pig BW and feed intakes were measured once daily throughout the experiment. Starting on d 1, the ghrelin pigs were intravenously infused three times daily for 5 d with 2 microg/kg BW of human ghrelin, and the control pigs were similarly infused with saline. Activity observations and blood samples were taken at -15, 0, 15, 30, 60, 90, 120, 240, and 480 min relative to the first infusion and then three times daily (0800, 1600, and 2400) for 8 d. Weight gain during the 5-d infusion period was greater by the ghrelin than by control pigs (0.57+/-0.10 vs. 0.21+/-0.13 kg, respectively; P < 0.04); however, there was no increase in feed intake. During two behavioral observation periods, more pigs in the ghrelin treatment were observed eating compared with control pigs (P < 0.05). The initial infusion of exogenous ghrelin increased serum ghrelin, GH, insulin, and cortisol concentrations (P < 0.05). Endogenous serum ghrelin increased from d 1 to 8 of the experiment in control animals (P < 0.05). Serum IGF-I initially fell in both treatment groups from d 1 to 2 (P < 0.05) but then increased from d 5 to 8 (P < 0.05). Peripheral concentrations of glucose in the ghrelin pigs were greater on d 2, 3, 7, and 8 than on d 1 (P < or = 0.05). In both treatment groups, peripheral concentrations of leptin increased from d 7 to 8, and cortisol decreased from d 1 to 5 of the experiment. These observations provide evidence that ghrelin may positively influence weight gain and concomitantly increase GH, insulin, and cortisol secretion in weaned pigs.     

Mannan oligosaccharide increases serum concentrations of antibodies and inflammatory mediators in weanling pigs experimentally infected with porcine reproductive and respiratory syndrome virus

Mannan-containing products are capable of modulating immune responses in animals. However, different products may have diverse immunomodulation. The experiment was conducted to examine effects of mannan oligosaccharide (Actigen; ACT) on growth performance and serum concentrations of antibodies and inflammatory mediators in weanling pigs (Sus scrofa) experimentally infected with porcine reproductive and respiratory syndrome virus (PRRSV). A total of 32 PRRSV-negative pigs (3 wk old) were randomly assigned from within blocks to 1 of 4 treatments in a 2 by 2 factorial arrangement [2 types of diet: control (0%) and ACT addition (0.04%); and with and without PRRSV] in a randomized complete block design. Pigs were blocked by initial BW within sex. Ancestry was equalized across treatments. Pigs (8/treatment) were kept individually in each pen. After 2 wk of an 8-wk period of feeding the treatments, pigs received an intranasal inoculation of PRRSV or sham medium at 5 wk of age. Infection by PRRSV decreased ADG, ADFI, and G:F throughout the experiment (P < 0.01). Actigen did not affect ADG (P = 0.450), but decreased (P = 0.047) ADFI from 28 to 42 days postinoculation (DPI). During that time, ACT improved G:F in infected pigs but not in sham controls (interaction, P = 0.009). Dietary ACT did not affect viremia in infected pigs (P > 0.05), but increased PRRSV-specific antibody titer at 35 DPI (P = 0.042). Infection with PRRSV induced the febrile responses of pigs from 3 to 10 DPI (P < 0.001) with return to normal at 14 DPI. During the experimental period, the rectal temperature of pigs was found slightly elevated by ACT (P = 0.045). Infected pigs had greater serum concentrations of IL-1β, tumor necrosis factor (TNF)-α, IL-12, interferon (IFN)-γ, IL-10, and haptoglobin (Hp) than sham controls (P < 0.001). These results indicate that PRRSV stimulated secretion of cytokines involved in innate, T-helper 1, and T-regulatory immune responses. Actigen tended to decrease the serum TNF-α concentration regardless of PRRSV (P = 0.058). The ACT × PRRSV interaction was significant for IL-1β (P = 0.016), IL-12 (P = 0.026), and Hp (P = 0.047), suggesting that infected pigs fed ACT had greater serum concentrations of these mediators than those fed the control. The increases in IL-1β and IL-12 may favorably promote innate and T-cell immune functions in infected pigs fed ACT. Feeding ACT may be useful as ACT is related to increased PRRSV antibody titers and G:F in infected pigs at certain times during infection.     

Dietary supplementation of a Saccharomyces cerevisiae fermentation product attenuates exercise-induced stress markers in young horses

Mitigation of exercise-induced stress is of key interest in determining ways to optimize performance horse health. To test the hypothesis that dietary supplementation of a Saccharomyces cerevisiae fermentation product would decrease markers of exercise-induced stress and inflammation in young horses, Quarter Horse yearlings (mean ± SD; 9 ± 1 mo) were randomly assigned to receive either no supplementation (CON; n = 8) or 21 g/d S. cerevisiae fermentation product (10.5 g/feeding twice daily; SCFP; n = 10) top-dressed on a basal diet of custom-formulated grain as well as ad libitum Coastal bermudagrass hay. After 8 wk of dietary treatments, horses underwent a 2-h submaximal exercise test (SET) on a free-stall mechanical exerciser. Serum was collected before dietary treatment supplementation (week 0), at week 8 pre-SET, and 0, 1, and 6 h post-SET and analyzed for concentrations of cortisol and serum amyloid A (SAA) by commercial enzyme-linked immunosorbent assay (ELISA) and for cytokine concentrations by commercial bead-based ELISA. Data were analyzed using linear models with repeated measures in SAS v9.4. From week 0 to 8 (pre-SET), serum cortisol decreased (P = 0.01) and SAA did not change, but neither were affected by diet. Serum concentrations of all cytokines decreased from week 0 to 8 (P ≤ 0.008), but granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor, and interleukin-8 (IL-8) decreased to a greater extent in CON than in SCFP horses (P ≤0.003). In response to the week 8 SET, serum cortisol increased in all horses (P < 0.0001) but returned to pre-SET levels by 1 h post-SET in horses receiving SCFP. At 6 h post-SET, cortisol concentrations in CON horses returned to pre-SET concentrations, whereas cortisol declined further in SCFP horses to below pre-SET levels (P = 0.0002) and lower than CON (P = 0.003) at that time point. SAA increased at 6 h post-SET in CON (P < 0.0001) but was unchanged through 6 h in SCFP horses. All cytokines except G-CSF increased in response to the SET (P < 0.0001) but showed differing response patterns. Concentrations of IL-1β, IL-6, and tumor necrosis factor-alpha were lesser (P ≤ 0.05), and concentrations of G-CSF and IL-18 tended to be lesser (P ≤ 0.09) in SCFP compared with CON horses throughout recovery from the SET. In summary, 8 wk of dietary supplementation with 21 g/d of SCFP may mitigate cellular stress following a single, prolonged submaximal exercise bout in young horses.     

Effects of mannan oligosaccharide on cytokine secretions by porcine alveolar macrophages and serum cytokine concentrations in nursery pigs

This study explored the hypothesis that mannan oligosaccharide (MOS) acts to reduce systemic inflammation in pigs by evaluating cytokine production of alveolar macrophages (AM) and serum cytokine concentrations. A total of 160 pigs were fed diets containing 0.2 or 0.4% MOS for 2 or 4 wk postweaning compared with control diets without MOS. Dietary MOS did not affect the serum concentration of tumor necrosis factor (TNF)-α and tended (P = 0.081) to increase that of IL-10. These cytokine concentrations also changed over time (P < 0.001). After 2-wk feeding of the control or MOS diets, AM were collected and stimulated ex vivo with lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (PLIC) as infection models. The LPS-stimulated AM from MOS-fed pigs (n = 12) secreted less TNF-α (P < 0.001) and more IL-10 (P = 0.026) than those from control-fed pigs (n = 6). However, dietary MOS had less effect on ex vivo TNF-α and IL-10 production by PLIC-stimulated AM (P = 0.091 and P > 0.10, respectively. Further, effects of MOS were examined in 4 in vitro experiments. In Exp. 1 (n = 4 pigs), MOS and mannan-rich fraction (MRF), when added to AM cultures, were able to increase TNF-α production. This direct effect of MOS was not due to endotoxin contamination as verified in Exp. 2 (n = 6 pigs) using polymyxin B, an inhibitor of LPS activation of toll-like receptor 4. Polymyxin B inhibited production of TNF-α by AM after treatment with LPS (P < 0.001), but not after treatment with MOS in the absence of LPS (P > 0.70). In Exp. 3 (n = 6 pigs), when MOS was directly applied in vitro, the pattern of cytokine production by LPS-activated AM was similar to that observed ex vivo, as MOS suppressed LPS-induced TNF-α (P < 0.001) and enhanced LPS-induced IL-10 (P = 0.028). In Exp. 4 (n = 6 pigs), when MRF replaced MOS, AM-produced TNF-α induced by LPS or PLIC was suppressed by MRF (P = 0.015 or P < 0.001, respectively). These data establish that MOS and MRF suppress LPS-induced TNF-α secretions by AM. Generally, the study suggests that MOS may be a potent immunomodulator because it directly activates AM to secrete TNF-α and alters the cytokine responses of bacterial endotoxin-induced AM in both ex vivo and in vitro systems. In particular, feeding MOS to pigs for 2 wk reduces TNF-α and increases IL-10 concentrations after ex vivo treatment of AM with LPS. These immunomodulatory properties of MOS may have important implications for both host defense and avoidance of harmful overstimulation of the immune system.