Response to adrenocorticotropic hormone or corticotrophin-releasing hormone and vasopressin in lactating cows fed an immunomodulatory supplement under thermoneutral or acute heat stress conditions.

Adrenal responsiveness was tested in nonpregnant, lactating Holstein dairy cows fed diets supplemented with OmniGen-AF (OG; Phibro Animal Health Corp., Teaneck, NJ), an immune modulator, and in nonsupplemented control (CON) cows following bolus infusions of a combination of corticotropin-releasing hormone (CRH; 0.3 µg/kg of BW) and arginine vasopressin (VP; 1.0 µg/kg of BW) or ACTH (0.1 IU/kg of BW) in 2 environments: thermoneutral [TN; temperature-humidity index (THI) <60] for 24 h/d and heat stress (HS; THI >68 for 17 h/d). Cows (506) were initially fed OG (n = 254) or CON (n = 252) diets for 44 d before selection of a subgroup of cows (n = 12; 6 OG, 6 CON) for the study. The 2 subgroups were balanced for parity, milk yield, and days in milk. All cows were transported to and housed in 2 environmentally controlled rooms at the University of Arizona Agricultural Research Complex (Tucson). Cows were given 3 d to acclimate to the rooms and then underwent 12 d of TN conditions and then 8 d of HS conditions for a total of 24 d on experiment. Cows were infused with CRH-VP on d 9 of TN and on d 1 of HS and with ACTH on d 10 of TN and on d 2 of HS. Hormone infusions took place at 1000 h (0 h) on each infusion day. Blood samples, taken in 30-min intervals, were first collected at 0800 h (-2 h) and were drawn until 1800 h (8 h). Before infusion, serum progesterone was elevated in OG cows compared with CON cows. Infusion of releasing factors (CRH-VP or ACTH) caused increases in serum cortisol and progesterone, but cortisol release was greater in CON cows than in OG cows during HS, whereas progesterone did not differ between the 2 treatments. Serum ACTH increased following infusion of releasing factors, but this increase was greater following CRH-VP infusion than ACTH infusion. Serum bovine corticosteroid-binding globulin also increased following infusion of releasing factors in both treatment groups, but this increase was greater during HS in cows fed OG. The free cortisol index (FCI) increased following CRH-VP and ACTH and was higher in HS than in TN for both OG and CON cows. However, the FCI response was blunted in OG cows compared with CON cows during HS. Heat stress enhanced the adrenal response to releasing factors. Additionally, the adrenal cortisol and FCI response to releasing factors was reduced during acute heat stress in cows fed OG. Collectively, these data suggest that OG supplementation reduced the adrenal responsiveness to factors regulating cortisol secretion during acute HS.

Relationships among temperament, acute and chronic cortisol and testosterone concentrations, and breeding soundness during performance testing of Angus bulls.

The aim of this study was to examine relationships among temperament, endocrinology, and reproductive parameters of bulls enrolled in an 84-day performance test. Angus bulls (n = 60) were housed in six pens grouped by age and weight. Pen scores (PS; 1 = docile to 5 = very aggressive) were assigned on Days -1, 27, 55, and 83 of the performance test. On the following day, blood and hair samples were collected, and body weight (BW) and exit velocity (EV) were recorded. Bulls were split into two categories based on; Day -1 PS (PScalm = PS 1 or 2; PSexcitable = PS 3 or 4) and Day 0 EV (EVcalm = slowest 20 bulls; EVexcitable = fastest 20 bulls). Cortisol and testosterone concentrations in serum and hair did not differ (P > 0.10) between PS or EV temperament categories. Sampling day differences (P < 0.01) occurred for serum testosterone, hair cortisol, and hair testosterone concentration; however, serum cortisol concentration did not differ (P > 0.10) over the sampling days. Serum testosterone concentration increased (P < 0.01) from Day 0 to 28, decreased from Day 28 to 56, but Day 84 did not differ from Day 0, 28, or 56. Hair cortisol concentration was greatest (P < 0.01) on Day 0, decreased from Day 28 to 56 but did not differ from Day 56 to 84. Hair testosterone concentration was greatest (P < 0.01) on Day 0 and remained constant from Day 28 to 84. Bulls categorized as PScalm had a greater (P < 0.01) percentage of normal sperm and secondary defects (P < 0.01) when compared with PSexcitable bulls. However, EVcalm bulls had fewer (P < 0.01) primary defects but more (P < 0.01) secondary defects than EVexcitable bulls. In conclusion, bulls exhibited physiological evidence of acclimation during the test as indicated by a reduction in hair cortisol concentration. In addition, the ability of the bulls to acclimate while residing at the testing center may have contributed to little differences observed during the breeding soundness examination portion of the performance test.

Relationships among temperament, behavior, and growth during performance testing of bulls.

Excitable cattle are dangerous to personnel and have reduced individual performance. The aim of this study was to 1) identify objective criteria for evaluating bull temperament and 2) examine relationships among temperament, behavior, and performance of bulls during an 84-d performance test. Angus bulls ( = 60) were reared in 6 pens based on BW and age. Pen scores (PS; 1 = docile and 5 = very aggressive) were assigned on d -1, 27, 55, and 83. Exit velocity (EV), BW, time to exit the chute, and order through the chute were recorded on d 0, 28, 56, and 84. The ADG was calculated for the 84-d test period, and ultrasound data and frame score calculations were recorded on d 84. Dataloggers measured steps taken, lying time, number of lying bouts, and lying bout duration of bulls ( = 27; 3 pens) from d 3 to 28 and d 59 to 84. Bulls with a d -1 PS of 1 or 2 were categorized as calm (PScalm; = 40), whereas bulls with a PS of 3 or 4 were categorized as excitable (PSexcitable; = 20). Bulls were separated into 2 groups based on the bottom 20 EV (EVcalm) and top 20 EV (EVexcitable) on d 0. Mixed model ANOVA (SAS 9.3) was used to compare groups for the two temperament assessment methods, behavior, and growth performance. Mean EV decreased ( < 0.05) by d 84. Total lying time from d 3 to 28 was greater ( < 0.05) for PScalm bulls when compared with PSexcitable bulls. However, total lying time from d 59 to 84 was greater ( < 0.05) for EVexcitable bulls when compared with EVcalm bulls. Regardless of initial contemporary group assignment, all bulls exited the chute slower ( < 0.001) on d 84 than on d 0. The PSexcitable bulls had greater ( < 0.01) frame scores and greater ADG than PScalm bulls. The PSexcitable bulls had more ( < 0.01) backfat than PScalm bulls. However, ribeye area was smaller ( < 0.01) in EVexcitable bulls than EVcalm bulls. Based on these results, bulls appeared to have habituated over the testing period. Additionally, the potential lack of innate temperament variation may have attributed to the little difference seen among the behavioral and performance data. Therefore, temperament should be reassessed within a novel environment with new handlers to differentiate between the bull's true temperament and its ability to habituate.

Effect of prenatal stress on subsequent response to mixing stress and a lipopolysaccharide challenge in pigs.

Sows subjected to prenatal stress have been found to produce offspring that have altered responses to stress. Our objective was to determine if exposing a sow to stress would alter the response of the offspring to lipopolysaccharide (LPS) at 2 mo of age or their response to mixing stress at 4 mo of age. Sow treatments consisted of intravenous injections of ACTH (1 IU/kg of BW), exposure to rough handling for a 10-min duration (rough), or no treatment (control) once per week from d 42 to 77 of gestation. At 2 mo of age, pigs from each treatment, 1 per litter (n = 21, 17, and 15 for the ACTH, rough, and control treatments, respectively), were challenged with 2 µg of LPS/kg of BW or saline, or served as a noninjected control. Their behavioral response to a human approach test and salivary cortisol were measured. At 4 mo of age, 1 pig from each treatment (n = 14, 14, and 15 for the ACTH, rough, and control treatments, respectively) was taken from its home pen and placed in a pen of unfamiliar pigs. At this time, a punch biopsy wound (6 × 6 mm) was created to measure the ability of the pig to heal the wound. At this same time, each pig received a 1-mL intramuscular injection of 20% ovine red blood cells (oRBC), and then a second injection of oRBC at 21 d postmixing. Blood samples were collected 3 times per week for 2 wk and then once a week for 4 more weeks. Blood samples were analyzed for cortisol, porcine corticosteroid-binding globulin, antibody response to oRBC, and nitric oxide production by macrophages. Behavior was recorded during the first 5 d after mixing. All pigs in the LPS challenge responded with characteristic sickness behavior; however, pigs in the rough treatment showed less sickness behavior than those in the other 2 treatments (P < 0.05). Maternal stress treatment did not affect (P < 0.43) salivary cortisol. Pigs from all treatments responded similarly to mixing stress with regard to cortisol, porcine corticosteroid-binding globulin, antibody titers, nitric oxide production, and hematology measures, and all pigs experienced the same amount of aggression in response to mixing. Without altering peripheral measures of stress responsivity, prenatal stress enhanced the ability of pigs to cope with a simulated immune challenge, which could prove to be an adaptation to challenging environments.

Effects of weaning and syndyphalin-33 on expression of melanocortinergic appetite-regulating genes in swine.

Syndyphalin-33 (SD-33) increases feed intake in sheep and recently weaned pigs. To assess the effects of SD-33 on hypothalamic gene expression, hypothalami were collected from unweaned pigs (n=19; 21±3 d of age) on day 0. Remaining pigs received an intramuscular injection of 0.5 µmole/kg SD-33 (SD) or saline (VEH) and weaned into individual pens. On days 1, 4, and 7 after weaning, hypothalami were collected from subsets of pigs (n=8 or 9) within each treatment group. Expression of µ-opioid receptor (MOR) was less in SD pigs than in VEH pigs on day 1 and day 4, suggesting down-regulation of the receptor by SD-33. Expression of hypothalamic melanocortin 4 receptor (MC4R) at 1 d after weaning was increased in VEH pigs (but not SD pigs) relative to levels before weaning. Expression of AGRP was not significantly altered by weaning or treatment at 1 d after weaning. At 4 d after weaning, expression of AGRP was greater in SD pigs than in VEH pigs, but at day 7 expression was less in SD pigs than in VEH pigs. A strong positive correlation was noted between expression levels of MOR and MC4R across treatment and time. Treatment with SD-33 appeared to partially abrogate the effects of weaning on expression of two key appetite-regulating genes within 24 h. Effects of SD-33 appear to be mediated at least in part by the µ-opioid receptor and include actions on the melanocortinergic pathway.

Effects of syndyphalin-33 on feed intake and circulating measures of growth hormone, cortisol, and immune cell populations in the recently weaned pig.

The synthetic met-enkephalin syndyphalin-33 (SD-33) increases feed intake in sheep and transiently increases circulating GH concentrations in sheep, rats, and pigs. Two experiments were performed to evaluate the effects of SD-33 on recently weaned pigs. In a preliminary experiment, pigs were administered SD-33 (0.5 micromol/kg, given intramuscularly) or saline immediately before a 3-h transport and subsequent placement into group pens. Treatment with SD-33 increased (P = 0.01) daily feed intake; cumulatively, pen intake over 7 d postweaning tended (P = 0.06) to be greater than in control pens. In Exp. 2, pigs were weaned and fitted with jugular catheters. The following day, pigs were treated with SD-33 or saline as described above. Transient increases (P < 0.05) in circulating concentrations of GH (at 1 and 1.5 h postinjection) and cortisol (at 3.5 and 4 h postinjection) were observed in pigs treated with SD-33 relative to controls. No difference in feed intake was observed between treatments over 4 d postinjection. Increased (P < 0.05) numbers of circulating neutrophils, lymphocytes, and monocytes were observed in both treatment groups over 4 d postinjection, and treatment with SD-33 tended (P = 0.07) to selectively increase monocyte numbers. Although SD-33 has potential to be used to increase feed intake and decrease the negative effects of stress during weaning in pigs, further investigation is needed to better understand the timing of effect and to rule out possible immunosuppressive effects.

Physiological and immunological responses to weaning and transport in the young pig: modulation by administration of porcine somatotropin.

To examine the effects of exogenous porcine (p) ST on measures of stress and immune function in weaned pigs with or without transport, pigs (20 +/- 1 d of age) received daily injections of pST (0.5 mg/kg; n = 16) or saline (n = 16) for 5 d. On d 5, a blood sample was collected immediately before injection. At 4 h postinjection, pigs were weighed, sampled for blood, injected with di-nitrophenyl-conjugated keyhole limpet hemocyanin, and weaned. One half of the pigs in each group were transported for 3 h before placement in the nursery. Pigs were weighed, and blood was collected on 1, 7, and 14 d postweaning. Statistical significance was set at P < 0.05. Serum IGF-I concentrations were increased by pST and decreased by weaning, but not affected by transport. The free cortisol index was elevated in all pigs 1 d postweaning, although less in transported versus nontransported pigs. By 7 d postweaning, the free cortisol index returned to prewean values. Serum concentrations of immunoglobulin (Ig) G increased in all pigs by 14 d postweaning, but were not affected by pST or transport. Serum IgM concentrations were elevated at 7 and 14 d postweaning. Before weaning and again 1 d postweaning, pigs treated with pST had greater concentrations of IgM than did control animals. Circulating neutrophils increased in pST-treated pigs 4 h after the final pST injection. Improved immune function in weaned pigs by pST may lead to greater health and growth in a commercial setting.

Prenatal stress effects on pig development and response to weaning.

Exposing a pregnant sow to stress has been shown to affect the resulting offspring. Our objective was to determine if rough handling of pregnant sows altered the physiology of her offspring and if these alterations were different from an experimentally induced model of prenatal stress. Sow treatments consisted of i.v. injections of ACTH (1 IU/kg of BW), exposure to rough handling for 10 min (Rough), or no treatment (Control) once a week during d 42 to 77 of gestation. To determine the plasma cortisol response to treatments, blood (5 mL) was collected from 30 sows after treatment administration. To conduct the prenatal stress study, a separate group of 56 sows was used in 1 of 4 replicates. At birth, production data were collected for each litter, including birth weight, number born, anogenital distance, and pig viability. At weaning, pigs were blocked by BW and sex, and placed in a nursery pen of 6 pigs, with 2 pigs from each treatment group. To assess the effect of treatments on cortisol, corticosteroid-binding globulin (CBG), and hematological cell profiles, blood was collected every other day for 10 d after weaning. Application of treatments caused plasma cortisol concentrations to be greatest in ACTH sows compared with Control sows (P < 0.001), with Rough sows having intermediate values (P = 0.07). Treatments did not affect the number of pigs born, number of stillborn, or pig viability (P > 0.40). The ratio of cortisol to CBG did not differ between treatments (P = 0.09). Hematological variables did not differ between treatments (P > 0.19). Pigs born to ACTH sows had a smaller anogenital distance compared with controls (P < 0.03), with pigs from Rough sows being intermediate. Our data indicate that swine exposed to prenatal stress (ACTH injection) can have alterations in sexual morphology without effects on growth or the immune cell populations measured in this study.

Effects of a concentrated lidocaine solution on the acute phase stress response to dehorning in dairy calves.

The objective of this study was to more fully define the surgical stress response to dehorning by heat cauterization in dairy calves by measuring behavioral, hormonal, inflammatory, and immunological markers of stress and to determine whether a nerve block of the surgical site with a concentrated solution of lidocaine (5%) reduces the degree of stress. Thirty-two 10- to 12-wk-old female Holstein calves were randomly allotted to 1 of 4 treatments: 5% lidocaine followed by dehorning, 2% lidocaine followed by dehorning, saline followed by dehorning, or 5% lidocaine followed by sham dehorning. Plasma cortisol concentration was measured in blood samples collected via a jugular catheter at -0.5, 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 9, 12, 24, 48, and 72 h. Various other blood constituents were measured in samples collected at -0.5, 12, 24, 48, and 72 h. Feeding, drinking, scratching, grooming, rubbing, licking, and inactivity behaviors were observed in the standing and recumbent positions using a 10-min scan sampling method analyzed on a time period and daily basis for 72 h following the dehorning procedure. The frequency of vocalization, kicking, and lying in the chute during the dehorning procedure were also assessed. The overall plasma cortisol concentrations were higher in calves subjected to dehorning than in control calves. Compared with the control group, the saline-treated calves had a higher cortisol concentration at 30 and 60 min postdehorning. Plasma cortisol concentrations were higher in all groups at 30 min postdehorning than at other sampling times. The percentage of circulating neutrophils and the neutrophil:lymphocyte ratio were increased in the saline and 2% lidocaine group. Total plasma protein, fibrinogen, and alpha1-acid glycoprotein concentrations were similar among treatments. The behavioral response to dehorning, as manifested by kicking while in the chute, was greater in the saline and 2% lidocaine group than in the control or 5% lidocaine treatment groups. In the postdehorning period, the percentage of time calves spent performing various maintenance behaviors did not differ among treatments. Thus, injection of 5% lidocaine may not provide any added comfort after the dehorning but may decrease the overall stress response during the procedure.

Temporal relationships between plasma cortisol, corticosteroid-binding globulin (CBG), and the free cortisol index (FCI) in pigs in response to adrenal stimulation or suppression.

The objective of this study was to document changes in plasma concentrations of total cortisol, porcine corticosteroid-binding globulin (pCBG), and the free cortisol index (FCI) in pigs over a 6-h period in response to adrenal stimulation or suppression. Twenty-four 8-week old pigs allotted in equal numbers were administered ACTH, dexamethasone or saline, and blood samples were collected every 15 min via an indwelling jugular catheter for 1 h prior to and 5 h following treatment. Total plasma cortisol increased in ACTH-treated pigs and decreased in dexamethasone-treated pigs within 0.25 and 0.5 h, respectively. In contrast, pCBG concentration was altered in an inverse fashion subsequent to the changes exhibited in total cortisol. FCI reflected the changes observed in total cortisol. These results further document the negative relationship that exists between circulating concentrations of plasma cortisol and pCBG, and illustrate that this association exists under conditions of acute stress in the pig.

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.

Hepatic corticosteroid-binding globulin (CBG) messenger RNA expression and plasma CBG concentrations in young pigs in response to heat and social stress.

Plasma cortisol, porcine corticosteroid-binding globulin (pCBG), hepatic CBG expression, and other physiological and behavioral measures of stress were studied in pigs in response to elevated temperature in conjunction with establishing a social hierarchy. Twenty-four crossbred pigs were weaned at 25 d of age (three or six pigs from six sows) and housed in littermate groups at 23 +/- 2 degrees C. At 57 d of age (d 0), animals were weighed and placed under general anesthesia for collection of blood (10 mL) and liver (approximately 100 mg) samples. On d 1, three unacquainted pigs of similar BW (23 +/- 1 kg) from different litters were allotted to each of eight nursery pens within two environmentally controlled rooms (12 pigs per room). From d 1 to 7, one room was maintained at 23 +/- 2 degrees C (CON) and the other at 33 +/- 2 degrees C (HEAT). Both rooms were kept at 23 +/- 2 degrees C from d 8 to 14. Animals were videotaped for 72 h beginning on d 1 and 8 to document behavioral changes in response to room temperature. The social hierarchy of pigs within each pen was based on fight activity recorded on d 1 to 3. Blood and liver tissue were collected again on d 7 and 14. The ADG for HEAT pigs increased (P < 0.05) over d 8 to 14 compared with d 1 to 7. In contrast to CON pigs, HEAT pigs displayed increased (P < 0.01) drinking but decreased feeding and lying in contact with other pigs from d 1 to 3, and similar drinking and feeding but increased (P < 0.01) lying with contact behaviors from d 8 to 10. With the exception of subordinate pigs exhibiting less (P < 0.05) frequent standing/walking behavior than the dominant or intermediate pigs on d 1 to 3, frequency of behaviors for both recorded time periods did not differ among pigs due to social status, regardless of treatment. The concentration of plasma haptoglobin in HEAT pigs on d 7 compared with d 0 increased (467 vs. 763 mg/L; P < 0.05), whereas cortisol and pCBG decreased (274 vs. 235 nmol/L and 11.4 vs. 9.9 mg/L, respectively; P < 0.05) as a result of treatment. The free cortisol index (total cortisol/pCBG) was greater (P < 0.05) in HEAT pigs on d 14 than on d 0 or 7. Hepatic CBG mRNA level was not affected by treatment. On d 14, HEAT pigs had plasma cortisol, pCBG, and haptoglobin concentrations similar to those of CON pigs. These results indicate that measured behavioral and physiological responses were not related to social status, and decreased circulating levels of cortisol and pCBG in pigs following a 7-d exposure to elevated temperature may not be determined by hepatic CBG mRNA expression.

Age-related changes in porcine corticosteroid-binding globulin (pCBG) as determined by an enzyme-linked immunosorbent assay.

The objectives of this study were to develop an assay for the direct measure of porcine corticosteroid-binding globulin (pCBG) and to confirm age-related changes in plasma pCBG concentration. Isolation and purification of pCBG from plasma was performed by affinity chromatography and HPLC-DEAE anion exchange techniques. Analysis by SDS-PAGE revealed two polypeptides (54 and 59 kDa) having similar amino acid homology (>50%) to previously reported sequences of seven mammalian species for the first 33 amino acids. Porcine CBG (20 ng/well) was immobilized to microtiter plates and standards or samples added along with rabbit antiserum developed against the purified pCBG. Goat anti-rabbit IgG-alkaline phosphatase conjugate was added followed by p-NPP substrate. The resultant color development was read at 405 nm. Intra- and interassay coefficients of variation (n=26) of a pooled sample were 10 and 15%, respectively. Age-related changes (P<0.001) in plasma pCBG concentration (n=203) from day 3 through 168 of age confirmed that, in the pig, changes seen in the percent distribution of cortisol among protein bound and free forms around day 28 of age are associated with an increase in CBG concentration.

Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. I. Effects on the immune axis when fed diets containing spray-dried plasma.

The objective of the present study was to evaluate the potential immunological benefit of adding menhaden fish oil to the diet of weaned pigs. Twenty-four crossbred male pigs were weaned at approximately 18 days of age and placed on a complex nursery diet containing 30% lactose and 7% plasma protein with 6% corn oil as the fat source (Cont, n=12) or with 5% menhaden fish oil and 1% corn oil as the fat source (MFO, n=12) for a period of 15 days. Body weights did not differ (P>0.78) between dietary groups either at the beginning or end of the 15 days feeding period. On day 15, all pigs were non-surgically fitted with an indwelling jugular catheter. On d 16, pigs received an i.v. injection of either saline (n=6/dietary group) or lipopolysaccharide (LPS; 150 microg/kg body weight; n=6/dietary group) and blood samples were collected at 30 min intervals for a period of 5h. Serum was harvested and stored at -80 degrees C for analysis of cortisol (CS), corticosteroid-binding globulin (CBG), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). There was no significant effect of diet on basal concentrations (Time 0) of any of the blood parameters analyzed. A Time x Treatment x Diet interaction (P<0.03) was observed for serum CS such that those pigs which consumed the MFO diet followed by LPS treatment had a reduced CS response as compared to the LPS-treated pigs on the Cont diet. A Time x Treatment interaction (P<0.01) was observed for serum CBG such that LPS treatment reduced circulating CBG as compared to the saline-treated pigs. Time x Treatment x Diet interactions were also observed for serum concentrations of TNF-alpha (P=0.084) and IFN-gamma (P=0.022) such that both the TNF-alpha and IFN-gamma response to the LPS challenge was lower in those pigs receiving the MFO diet as compared to the LPS-treated pigs on the Cont diet. Overall, serum CS was negatively correlated with the CBG response (r=-0.40, P<0.001), however, the strongest negative correlation was observed in the LPS-treated pigs which consumed the MFO diet (r=-0.63, P<0.001). While further studies are needed to evaluate the immunological response of including MFO in the nursery pig diet, the present study demonstrates that supplementation with MFO does indeed alter the immunological response to an LPS challenge.

Partial nucleotide sequence of the porcine corticosteroid-binding globulin (CBG) cDNA and specification of CBG expression sites in postnatal pigs.

The purpose of this study was to develop a porcine CBG cDNA probe in order to examine the porcine CBG mRNA expression in major tissues from the postnatal pig. The reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to develop the porcine CBG cDNA probe using total RNA extracted from liver of 40-day-old pig. The RT-PCR product was subcloned into the pGEM vector (Promega, Madison, WI) and subjected to restriction enzyme treatments and DNA sequencing. Northern blot analysis was conducted using total RNA extracted from samples (approximately 200 mg) of liver, lung, kidney, and whole adrenal tissue that were collected from pigs on day 3 (n = 2) or day 40 (n = 2) postpartum. A 500 bp partial porcine CBG cDNA encoded 166 amino acids and had 83, 78, and 77% homology to a 494-nucleotide sequence of CBG from sheep, human, and rabbit, respectively. The deduced peptide sequence of the partial porcine CBG showed 77, 62, 60, and 51% homology to sheep, human, rabbit, and rat CBG sequences, respectively. An approximately 1.53 kb CBG mRNA was detected only in the liver tissue. In conclusion, the development of a partial CBG cDNA for swine makes it possible to study the ontogeny and the regulation of CBG synthesis at the molecular level and, based on tissues examined in this study, the liver appears to be the primary source of CBG biosynthesis in the postnatal pig.

The effects of split marketing on the physiology, behavior, and performance of finishing swine.

One hundred twenty 8-wk-old barrows (20.3 +/- 2.0 kg BW) were used to examine the effect of split marketing on selected behavioral, physiological and performance parameters. Pigs were assigned by weight in a randomized complete block design to one of three treatments: SM (split-marketed), six pigs/pen (1.83 m2/pig); C (control), six pigs/pen (1.83 m2/pig); or MC (modified control), three pigs/pen (3.66 m2/pig). The heaviest half of SM animals were removed 1 wk prior to marketing penmates. Control and MC animals remained in their respective groups until marketing. Animals were videotaped during the first 72 h of the study (INITIAL), 72 h prior to (PRE), and following the removal (POST) of pigs in the SM treatment to quantify maintenance behaviors and to identify socially dominant, intermediate, and submissive pigs. A blood sample was collected from each animal upon completion of INITIAL, PRE, and POST time periods to determine neutrophil:lymphocyte ratio and plasma haptoglobin, cortisol, and corticosteroid-binding globulin (CBG) levels. Animals were weighed and feed disappearance was calculated biweekly. Tenth-rib backfat and area of the longissimus muscle at marketing were ultrasonically evaluated on all animals. Regardless of treatment, animals were more (P < 0.01) active (eating, standing/walking, fighting) at INITIAL than at PRE or POST times. Frequency and duration of fights per pen were less (P < 0.01) in MC than in C or SM pigs for all periods observed. Neutrophil:lymphocyte ratio, plasma haptoglobin, and CBG levels were greater (P < 0.01) during the INITIAL period than during the PRE or POST periods but did not differ between treatments. No treatment or time differences were detected in plasma cortisol levels. The MC pigs exhibited greater (P < 0.01) ADFI with poorer feed efficiency compared to C or SM pigs up to split marketing. During the POST period, both MC and SM pigs had greater (P < 0.01) ADFI with poorer (P < 0.01) feed efficiency than C pigs. The ADG was not different among animals as a result of treatment. There were no treatment differences for any of the carcass measurements. Significant differences in performance between the treatment groups could not be attributed to any physiological or behavioral measures reported here.

Few differences found between early- and late-weaned pigs raised in the same environment.

Segregation and medicated early weaning are technologies used to optimize the productivity and health of pigs, but these practices may also cause aberrant behaviors indicative of stress. Thus, differences in early- (=10 d of age) and late- (=30 d of age) weaned pigs were investigated. At weaning, pigs were housed in groups of four in 16 pens (eight pens per treatment) in the same facility, and, thus, they were not segregated. Body weights were recorded at birth, weaning, and at approximately 42, 65, 102, 137, and 165 d of age (at slaughter). One-minute, instantaneous scan samples during a 10-min period (at 0600, 1000, 1400, and 1800) were used to record the frequency of lying, standing, and sitting, total number of drinks, feeder investigations, and time spent playing/fighting on 2, 3, and 4 d after weaning. Five-minute, direct observations of each pig were conducted at approximately 40, 60, 80, and 150 d of age. Direct observations were also made of the entire pen for 10 min at approximately 50, 95, 123, and 160 d of age to record aberrant behaviors. At 62 d of age, a handling and blood collection stress was imposed. At 165 d of age, a second stress test was conducted in response to rough handling and transport. Early-weaned pigs spent more time playing/ fighting (P < .006) than late-weaned pigs during the 4 d after weaning, manipulated conspecifics more often at 40 d of age (P < .002), had greater percentage of hemoglobin (P < .03) during Stress Test 1, had greater ADG at 42 d of age (P < .03), and had greater hypothalamic growth hormone-releasing hormone receptor mRNA at slaughter (P < .06). Late-weaned pigs had greater ADG between 137 and 165 d of age (P < .03) and greater pro-opiomelanocortin at slaughter (P < .04). Overall, most differences found between early-weaned and late-weaned pigs were evident soon after weaning, but they disappeared before slaughter.

Behavioral, endocrine, immune, and performance measures for pigs exposed to acute stress.

Weanling pigs (n = 132) were used to investigate the effects of three common stressors (and a control) and differing social status on behavior, immunity, plasma cortisol, blood chemical, and performance measures. Eleven blocks of 12 pigs each were evaluated. Each pen contained three pigs of dominant (DOM), intermediate (INT), or submissive (SUB) social status. Two weeks later, random pens of pigs experienced either a control treatment (CON) or they were stressed for 4 h by shipping (SHIP), heat-stressed (HEAT) with overhead heat lamps in their home pens, or cold-stressed (COLD) by direct application of water and an air current. Treatments did not influence body weights; however, percentage weight loss during SHIP was greater than for other treatments. Body weights were heavier for DOM pigs than for INT and SUB pigs. Social status had large effects on plasma cortisol, globulin, acute-phase proteins, body weight, and weight changes. Only acute shipping stress resulted in weight loss. Many immune and blood measures were not changed among acutely stressed pigs; however, the relationship between social status and mitogen-induced lymphocyte proliferation and natural killer cell cytotoxicity was disrupted during acute stress. Pig behavior was significantly changed by each stress treatment in a unique manner. During acute stress, behavioral changes seem to be the most consistent and reliable indicators.

Intrauterine cortisol, aldosterone, and corticosteroid binding globulin-like activity during early porcine pregnancy and the estrous cycle.

Studies were conducted to determine whether the corticosteroids cortisol and aldosterone, and corticosteroid-binding globulin (CBG) were present in the porcine early-embryonic environment. Cortisol was measured in uterine flushings from white crossbred gilts at Days 7, 10, 13, and 16 of the estrous cycle and pregnancy. Total content of cortisol increased (p < 0.01) between Days 13 and 16, and immunoreactive CBG (ir-CBG) increased (p < 0.01) between Days 10 and 13, in both cyclic and pregnant gilts. In a separate study with Chinese Meishan gilts, total cortisol and aldosterone content of uterine flushings increased (p < 0.02) between Days 10 and 15 of the estrous cycle and pregnancy. In another study with white crossbred gilts, CBG-like binding activity in uterine flushings was low at Day 10, then increased over 100-fold at Day 15 (p < 0.01). However, levels of CBG-like binding activity on Day 15 were 100-fold lower than those of ir-CBG measured in the previous study and could bind less than 4% of the uterine luminal cortisol. Differences between ir-CBG and CBG binding might be due to the ability of the CBG antibody to recognize either biologically inactive CBG or structurally similar molecules. CBG-like binding activity, which appeared unrelated to glucocorticoid receptors, was also present in the endometrial cytosol of white crossbred gilts. Concentrations (fmol/mg protein) of endometrial CBG-like activity decreased (p = 0.03) between Days 10 and 15 of the estrous cycle and pregnancy, did not differ with reproductive status, and on Day 15 were comparable to concentrations in uterine flushings but threefold lower (p < 0.01) than those in the serum. Equilibrium dissociation constants for CBG-like binding activities were comparable among the three locations. These studies indicate that corticosteroids are present-primarily in the free form-within the porcine uterine lumen and could influence early porcine conceptus development. Endometrial CBG-like binding activity could mediate actions of cortisol or progesterone on uterine function.

Distribution between protein-bound and free forms of plasma cortisol in the gilt and fetal pig near term.

Thirty-five time-dated pregnant gilts were used to document plasma levels of total and free cortisol, corticosteroid-binding globulin (CBG) binding capacity, and percent distribution of cortisol among protein-bound (CBG and albumin) and free forms in the fetal pig during the last 24 days of gestation. Plasma from fetal pigs on days 110-114 of gestation (gestation length 114 days) had significantly higher levels of total cortisol (p < 0.01), percent albumin-bound and free cortisol (p < 0.10), and free cortisol concentration (p < 0.05) compared to samples on days 90, 100 and 105. Fetal plasma CBG binding capacity increased (p < 0.05) linearly from day 100 to 114. Fetal pigs located in the cervical region of the uterus had lower (p < 0.05) total and free cortisol and higher (p < 0.05) albumin and total protein concentrations compared to fetuses in the middle and oviductal regions. Total, percent free and free cortisol concentrations in maternal plasma on days 105-114 were greater (p < 0.10) than that measured on days 12-100 of gestation. These results suggest that the developmental patterns of plasma cortisol and CBG in the prenatal pig are directly related and highly similar to those of another precocious species, the sheep.