Investigating the effects of jute nesting material and enriched piglet mats on sow welfare and piglet survival.

Domesticated sows are motivated to perform nesting behavior prior to farrowing, and nesting material can reduce piglet cold stress. However, nesting material may not be practical in most production systems due to the potential for clogging slurry systems. Therefore, the study objectives were to assess an alternative nesting material provided prior to farrowing on sow welfare and piglet survival, and to investigate the effect of the entire nesting environment on piglet survival and growth performance. We hypothesized that the provision of jute nesting material would decrease sow stress and farrowing duration, and that nesting mat provision would allow piglets to remain euthermic and improve survival and growth. Sows (N = 20) were randomly assigned to one of two treatments: a farrowing crate with three pieces of 40.6 × 21.6 cm jute nesting material (Nest; n = 10) and two enriched piglet mats made from an acrylic board (28.0 × 86.4 cm) covered with a microfiber material, or a farrowing crate without nesting material (Control; n = 10) and one standard plastic piglet mat (28.0 × 86.4 cm). Jute pieces were attached to the front of the crate to prevent substrate from falling through the slatted floors. Saliva samples were collected to measure cortisol and immunoglobulin A (IgA), on days -1, 0, 1, and 2 relative to farrowing, and a final sample was collected at weaning (day 16.9 ±â€…0.18). Blood was collected from four piglets per litter to measure immunoglobulin G (IgG) at 48 h, day 7, and weaning. Piglet skin temperature (TS) was measured on two piglets per litter using an infrared camera for 3 d after birth at 0800, 1200, 1600, and 2000 h. One piglet was randomly chosen from the heat lamp and nonheat lamp side of the crate to measure TS. Video was continuously coded for observations of jute- and crate-directed interactions. Data were analyzed as a mixed model analysis of variance in SAS 9.4. Nest sows performed less crate-directed behavior than Control sows (P = 0.02). Cortisol tended to be reduced in Nest sows (P = 0.08) when compared to Controls, but no differences in IgA concentrations (P > 0.40) were detected. Nest piglets tended to be heavier on day 7 (P < 0.10), had greater IgG concentrations (P = 0.03), and had greater TS (P = 0.02) versus Controls. No farrowing duration or number of stillbirth differences were observed (P > 0.70). The jute material and piglet nests positively impacted sow welfare and piglet measures but did not translate into improved piglet survival.

Interactions between corticotropin releasing factor signaling and prophylactic antibiotics on measures of intestinal function in weaned and transported pigs.

The study objective was to evaluate the interaction between corticotrophin releasing factor (CRF) receptor signaling and prophylactic antibiotic administration on intestinal physiology in newly weaned and transported pigs. Pigs (n = 56; 5.70 ± 1.05 kg) were weaned (20.49 ± 0.64 d), a blood sample was taken, and then pigs were given an intraperitoneal injection of saline (SAL; n = 28 pigs) or a CRF receptor antagonist (CRFA; n = 28 pigs; 30 µg/kg body weight; Astressin B), and then were transported in a livestock trailer for 12 h and 49 min. A second and third intraperitoneal injection was given at 4 h 42 min and 11 h 36 min into the transport process, respectively. Following transport, 4 SAL and 4 CRFA pigs were blood sampled and euthanized. The remaining 48 pigs were individually housed and given dietary antibiotics [AB; n = 12 SAL and 12 CRFA pigs; chlortetracycline (441 ppm) + tiamulin (38.6 ppm)] or no dietary antibiotics (NAB; n = 12 SAL and 12 CRFA pigs) for 14 d post-transport. Blood was collected at 12 h and on d 3, 7, and 14, and then pigs were euthanized on d 7 (n = 24) and d 14 (n = 24) post-weaning and transport. Circulating cortisol was reduced (p = 0.05) in CRFA pigs when compared to SAL pigs post-weaning and transport. On d 7, jejunal villus height and crypt depth was greater overall (p < 0.05) in AB-fed pigs versus NAB-fed pigs. On d 14, ileal crypt depth was reduced (p = 0.02) in CRFA pigs when compared to SAL pigs. Jejunal CRF mRNA abundance tended to be reduced (p = 0.09) on d 7 in CRFA pigs versus SAL pigs. On d 14, jejunal tumor necrosis factor-alpha was reduced (p = 0.01) in AB-fed pigs versus NAB-fed pigs. On d 7, change in glucose short-circuit current tended to be increased (p = 0.07) in CRFA pigs fed the AB diet when compared to CRFA pigs fed the NAB diet. In conclusion, CRFA pigs and pigs fed AB had some similar biological intestinal function measures post-weaning and transport.

Characterizing the effect of incrementally increasing dry bulb temperature on linear and nonlinear measures of heart rate variability in nonpregnant, mid-gestation, and late-gestation sows.

Characterizing the sow physiological response to an increased heat load is essential for effective heat stress mitigation. The study objective was to characterize the effects of a 400-min heating episode on sow heart rate variability (HRV) at different reproductive stages. HRV is a commonly used noninvasive proxy measure of autonomic function. Twenty-seven sows were enrolled in the study according to their gestation stage at time of selection: 1) nonpregnant (NP; n = 7), 2) mid-gestation (MID; 57.3 ± 11.8 d gestation; n = 11), and 3) late-gestation (LATE; 98.8 ± 4.9 d gestation; n = 8). The HRV data utilized in the study were collected from each pig as the dry bulb temperature in the room increased incrementally from 19.84 ± 2.15 °C to 35.54 ± 0.43 °C (range: 17.1-37.5 °C) over a 400-min period. After data collection, one 5-min set of continuous heart rate data were identified per pig for each of nine temperature intervals (19-20.99, 21-22.99, 23-24.99, 25-26.99, 27-28.99, 29-30.99, 31-32.99, 33-34.99, and 35-36.99 °C). Mean inter-beat interval length (RR), standard deviation of r-r intervals (SDNN), root mean square of successive differences (RMSSD), high frequency spectral power (HF), sample entropy (SampEn), short-term detrended fluctuation analysis (DFAα1), and three measures (%REC, DET, LMEAN) derived from recurrence quantification analysis were calculated for each data set. All data were analyzed using the PROC GLIMMIX procedure in SAS 9.4. Overall, LATE sows exhibited lower RR than NP sows (P < 0.01). The standard deviation of r-r intervals and RMSSD differed between each group (P < 0.01), with LATE sows exhibiting the lowest SDNN and RMSSD and NP sows exhibiting the greatest SDNN and RMSSD. Late-gestation sows exhibited lower HF than both MID and NP sows (P < 0.0001), greater DFA values than NP sows (P = 0.05), and greater DET compared to MID sows (P = 0.001). Late-gestation also sows exhibited greater %REC and LMEAN compared to MID (P < 0.01) and NP sows (all P < 0.01). In conclusion, LATE sows exhibited indicators of greater autonomic stress throughout the heating period compared to MID and NP sows. However, temperature by treatment interactions were not detected as dry bulb increased. Future studies are needed to fully elucidate the effect of gestational stage and increasing dry bulb temperature on sow HRV.

Pregnant pigs may be at a higher risk of poor physiological outcomes due to heat exposure compared to mature female pigs that are not pregnant. The purpose of this study was to evaluate the stress response of pregnant pigs to increasing environmental temperatures using heart rate variability, a noninvasive measure commonly used to evaluate the physiological stress response. Our findings show that pregnant pigs, particularly those who are closer to giving birth, exhibit greater evidence of physiological stress compared to pigs who are not pregnant. However, we did not find evidence that increasing environmental temperature throughout the experimental period was a primary reason for the increased stress exhibited by pregnant pigs. It is possible that the physiological changes that normally occur during pregnancy may have masked the physiological stress response typically associated with increased heat exposure.

A behavior and physiology-based decision support tool to predict thermal comfort and stress in non-pregnant, mid-gestation, and late-gestation sows.

BACKGROUND: Although thermal indices have been proposed for swine, none to our knowledge differentiate by reproductive stage or predict thermal comfort using behavioral and physiological data. The study objective was to develop a behavior and physiology-based decision support tool to predict thermal comfort and stress in multiparous (3.28 ± 0.81) non-pregnant (n = 11), mid-gestation (n = 13), and late-gestation (n = 12) sows. RESULTS: Regression analyses were performed using PROC MIXED in SAS 9.4 to determine the optimal environmental indicator [dry bulb temperature (TDB) and dew point] of heat stress (HS) in non-pregnant, mid-gestation, and late-gestation sows with respiration rate (RR) and body temperature (TB) successively used as the dependent variable in a cubic function. A linear relationship was observed for skin temperature (TS) indicating that TDB rather than the sow HS response impacted TS and so TS was excluded from further analyses. Reproductive stage was significant for all analyses (P < 0.05). Heat stress thresholds for each reproductive stage were calculated using the inflections points of RR for mild HS and TB for moderate and severe HS. Mild HS inflection points differed for non-pregnant, mid-gestation, and late gestation sows and occurred at 25.5, 25.1, and 24.0 °C, respectively. Moderate HS inflection points differed for non-pregnant, mid-gestation, and late gestation sows and occurred at 28.1, 27.8, and 25.5 °C, respectively. Severe HS inflection points were similar for non-pregnant and mid-gestation sows (32.9 °C) but differed for late-gestation sows (30.8 °C). These data were integrated with previously collected behavioral thermal preference data to estimate the TDB that non-pregnant, mid-gestation, and late-gestation sows found to be cool (TDB < TDB preference range), comfortable (TDB = TDB preference range), and warm (TDB preference range < TDB < mild HS). CONCLUSIONS: The results of this study provide valuable information about thermal comfort and thermal stress thresholds in sows at three reproductive stages. The development of a behavior and physiology-based decision support tool to predict thermal comfort and stress in non-pregnant, mid-gestation, and late-gestation sows is expected to provide swine producers with a more accurate means of managing sow environments.

Immunological, physiological, and behavioral effects of Salmonella enterica carriage and shedding in experimentally infected finishing pigs.

Finishing pigs infected with Salmonella pose significant food safety risks by carrying the pathogen into abattoirs. This study was conducted to determine the dynamics of Salmonella infection in finishing pigs, and associated immunological, physiological, and behavioral alterations, by longitudinally comparing infected to noninfected pigs during 6 weeks postinfection (p.i.). Bacteriological data revealed that all inoculated pigs started shedding Salmonella within 2 h p.i., and persistently shed the bacteria up to the end of the study. Ileal and cecal contents, as well as mesenteric lymph node samples, were all positive throughout the study, containing 3-4 log(10) cfu/g of Salmonella at 24 h p.i., and 4-5 log(10) cfu/g of Salmonella up to 4 weeks p.i. Levels of Salmonella dropped markedly (p < 0.05) in all samples at 5 weeks p.i. There was no difference between groups for blood cell counts. Tumor necrosis factor-α was greater (p < 0.05) in infected pigs: (1) in the mesenteric lymph nodes by 48 h p.i.; (2) at 24 h and 3 weeks p.i. in the ileum; and (3) in the cecum and spleen at 3 weeks p.i. Interleukin-12, interleukin-1 and its antagonist, and a porcine-specific antimicrobial peptide RNA expression in tissues changed over time, but were not different between groups. Infected pigs spent more time in ventral recumbency, standing, and sitting than controls (p < 0.01). Infected pigs were also more active (p < 0.01), and approached a novel object more quickly than control pigs (p < 0.05). No treatment differences were detected for rectal temperature or plasma cortisol (p > 0.10). This study shows that finishing pigs can carry high levels of Salmonella for up to 4 weeks p.i. in the gastrointestinal contents and mesenteric lymph nodes, shedding high levels of the bacteria without developing clinical symptoms, but developing an immune response throughout the intestinal tract. Moreover, subtle behavioral changes measured as postures were detected, and therefore warrant additional investigation.

Effect of short-term high tryptophan diet fed to sows on their subsequent piglet behavior.

Housing sows in groups create the challenge of decreasing fighting amongst sows. One proposed method to do so is to feed a high tryptophan diet, but the effect on the fetus is unknown. To investigate this, 66 sows were fed one of three diets: Control (0.14% SID tryptophan), Medium (0.28% SID tryptophan), or High (0.42% SID tryptophan), from days 28 to 35 of gestation. Sows gestated in standard gestation stalls. Blood samples were taken on day 27 prior to and on day 35 after tryptophan supplementation. On days 1, 2, and 3, nursing bouts were observed so as to record disputes and displacements from teat competition. The piglets' activity and fighting were recorded on days 3, 7, and 11 from 0700 to 1700 h. On day 12, four piglets per litter were blood sampled: two to be used in later behavior tests and two to act as controls for blood cortisol levels. On day 14, the two behavior test piglets from each litter were subjected to a 10-min Isolation Test and 5-min Human Approach Test. On day 15, the behavior test piglets were paired by sex and treatment (for example, a male Medium piglet paired with another male Medium piglet from a different crate) and each pair was subjected to a 10-min Social Challenge Test and immediately blood sampled. Piglet cortisol and serotonin did not differ among treatments (P > 0.10). There were no differences (P > 0.10) for number born (12.7 ± 0.4), born alive (11.7 ± 0.4), or mortality (1.1 ± 0.2). Behavior during nursing bouts was similar, with no treatment differences in number of disputes or displacements, and similar bout lengths among treatments (199.5 ± 4.6 s, P > 0.10). No differences were detected for any of the variables for Isolation or the Human Approach Tests (P > 0.10). During the Social Challenge Test, High piglets had more contacts approaching the head of the companion piglet than did either Medium or Control piglets (14.3 ± 1.1, 10.7 ± 1.1, and 9.69 ± 0.8, respectively, P < 0.02). Total number of aggressive interactions during the test tended to be greater for Medium piglets compared to High piglets (9.3 ± 1.5 vs 5.1 ± 0.9, P < 0.07). Time budget data of the litter indicate that piglets from all three treatments spent equal amounts of time active and inactive (P > 0.10). Aggression was low with 0.3 ± 0.04% of piglets displaying aggressive behavior. Feeding high concentrations of tryptophan for a short duration early in gestation does not have a negative impact on sows' subsequent offspring.

Thermoregulatory and physiological responses of nonpregnant, mid-gestation, and late-gestation sows exposed to incrementally increasing dry bulb temperature.

Gestating sows may be more susceptible to increasing dry bulb temperatures (TDB) due to greater metabolic heat production and increased body mass, especially as gestation advances. However, there are few studies on the thermoregulatory and physiological responses of sows at differing gestation stages exposed to gradually increasing temperatures. The study objective was to determine the thermoregulatory and physiological responses of nonpregnant (n = 12; parity 3.27 ± 0.86), mid-gestation (59.7 ± 9.6 d pregnant, n = 12; parity 3.25 ± 0.83), and late-gestation (99.0 ± 4.8 d pregnant, n = 12; parity 3.33 ± 0.75) sows exposed to increasing TDB. Prior to the experiment (5.0 ± 0.7 d), jugular catheters were placed in all sows. During the experiment, the TDB was increased incrementally by 2.45 ± 0.43 °C every 60 min from 19.84 ± 2.15 to 35.54 ± 0.43 °C over 400 min, and relative humidity was recorded at 40.49 ± 18.57%. Respiration rate (RR), heart rate (HR), skin temperature, and vaginal temperature were measured, and blood samples were obtained via the jugular catheter every 20 min. Data were analyzed using PROC MIXED in SAS 9.4. RR increased at a lower TDB (P < 0.01) in late-gestation sows compared with mid-gestation and nonpregnant sows, but no differences were detected between mid-gestation and nonpregnant sows. Overall, late-gestation sows had greater RR (P < 0.01; 23 ± 2 breaths per min [brpm]) compared with mid-gestation (16 ± 2 brpm) and nonpregnant (15 ± 2 brpm) sows. Late-gestation sows had an overall greater HR (P < 0.01; 84 ± 5 beats per min [bpm]) than mid-gestation (76 ± 5 bpm) and nonpregnant (69 ± 5 bpm) sows. Late-gestation sows had overall reduced bicarbonate and total carbon dioxide levels (P = 0.02; 23.89 ± 1.97 and 25.41 ± 2.07 mmol/L, respectively) compared with mid-gestation (27.03 ± 1.97 and 28.58 ± 2.07 mmol/L, respectively) and nonpregnant (26.08 ± 1.97 and 27.58 ± 2.07 mmol/L, respectively) sows. Moreover, late-gestation sows had overall greater nitric oxide levels (P < 0.01; 248.82 ± 34.54 µM) compared with mid-gestation (110.47 ± 34.54 µM) and nonpregnant (41.55 ± 34.54 µM) sows. In summary, late-gestation sows appear to be more sensitive to increasing TDB as indicated by thermoregulatory and physiological responses when compared with mid-gestation or nonpregnant sows. The results from this study provide valuable information regarding thermoregulatory thresholds of sows at differing gestation stages.

Evaluation of sow thermal preference across three stages of reproduction.

The metabolic heat production of modern pigs has increased by an average of 16%, compared with sows of 30 years ago. Therefore, it is likely that temperature recommendations require updating to meet the needs of modern pigs. The objective of this study was to evaluate whether different reproductive stages of sows altered thermal preference and if current recommendations required updating. Twenty multiparous sows (3.4 ± 1.2 parity) in different reproductive stages (nonpregnant: n = 7; mid-gestation: 58.5 ± 5.68 d, n = 6; and late-gestation: 104.7 ± 2.8 d, n = 7) were tested. Thermal preference was individually tested, and sows could freely choose a temperature, using a thermal gradient between 10.4 and 30.5 °C. Sows were given 24 h to acclimate to the thermal apparatus. Before testing began, sows were given daily feed allotment and returned to the apparatus. Video from the 24-h test period was used to record sow behavior (time spent inactive), posture (upright and sternal and lateral lying), and location using instantaneous scan samples every 15 min. Data were analyzed using PROC MIXED procedure in SAS 9.4. A cubic regression model was used to calculate the sow's most preferred temperature based on the location, or temperature, in which they spent the most time. The preference range was calculated using peak temperature preference ±SE for each sow. The reproductive stage altered where sows spent their time within the thermal gradient (P < 0.01). Late-gestation sows preferred cooler temperatures (14.0 °C) than mid-gestation (14.8 °C; P < 0.01) and nonpregnant sows (14.8 °C; P < 0.01). In summary, sow thermal preferences were within the lower half of the current recommended range (10 to 25 °C). This indicates that temperatures at the higher end of the recommended range could be uncomfortable to sows and that the thermal comfort zone of sows may be narrower than recommendations indicate.

Replacing dietary antibiotics with 0.20% l-glutamine in swine nursery diets: impact on intestinal physiology and the microbiome following weaning and transport.

Previous research demonstrates that supplementing 0.20% l-glutamine (GLN) in the diets of newly weaned and transported pigs improves growth rate to a similar extent as providing dietary antibiotics (AB). However, research comparing the effects of GLN vs. AB on intestinal physiology and the microbiome is limited. Therefore, the study objective was to compare the effects of supplementing nursery diets with GLN, AB, or no dietary antibiotics (NA) on intestinal physiology and the microbiome of pigs in a production environment following weaning and transport. Mixed-sex piglets (N = 480; 5.62 ± 0.06 kg body weight [BW]) were weaned (18.4 ± 0.2 d of age) and transported for 12 h in central Indiana, for two replicates, during the summer of 2016 and the spring of 2017. Pens were blocked by BW and allotted to one of the three dietary treatments (n = 10 pens/dietary treatment/replicate [8 pigs/pen]): AB (chlortetracycline [441 ppm] + tiamulin [38.6 ppm]), GLN (0.20% as-fed), or NA fed for 14 d. From day 14 to 34, pigs were fed common AB-free diets in two phases. On day 33, villus height:crypt depth tended to be increased (P = 0.07; 7.0%) in GLN and AB pigs vs. NA pigs. On day 33, glucagon-like peptide 2 (GLP-2) mRNA abundance was decreased (P = 0.01; 50.3%) in GLN and NA pigs vs. AB pigs. Crypt depth was increased overall on day 33 (P = 0.01; 16.2%) during the spring replicate compared with the summer replicate. Villus height:crypt depth was reduced (P = 0.01; 9.6%) during the spring replicate compared with the summer replicate on day 33. On day 13, tumor necrosis factor-alpha and occludin mRNA abundance was increased (P ≤ 0.04; 45.9% and 106.5%, respectively) and zonula occludens-1 mRNA abundance tended to be greater (P = 0.10; 19.2%) in the spring replicate compared with the summer replicate. In addition, AB pigs had increased (P = 0.01; 101.3%) GLP-2 mRNA abundance compared with GLN and NA pigs. Microbiome analysis indicated that on day 13, dietary treatment altered the microbiota community structure (P = 0.03). Specifically, the AB pigs tended to be distinct from both the NA and GLN pigs (P = 0.08), and Lactobacillus was increased nearly 2-fold in AB compared with NA pigs (q = 0.04) and GLN pigs (q = 0.22). In conclusion, GLN supplementation tended to improve some morphological markers of intestinal health similarly to AB pigs, while the microbiome composition in GLN pigs was more similar to NA pigs than AB pigs.

Characterizing the postnatal hypothalamic-pituitary-adrenal axis response of in utero heat stressed pigs at 10 and 15 weeks of age.

In utero heat stress alters postnatal physiological and behavioral stress responses in pigs. However, the mechanisms underlying these alterations have not been determined. The study objective was to characterize the postnatal hypothalamic-pituitary-adrenal axis response of in utero heat-stressed pigs. Pigs were subjected to a dexamethasone suppression test followed by a corticotrophin releasing hormone challenge at 10 and 15 weeks of age. Following the challenge, hypothalamic, pituitary, and adrenal tissues were collected from all pigs for mRNA abundance analyses. At 10 weeks of age, in utero heat-stressed pigs had a reduced (P < 0.05) cortisol response to the corticotrophin releasing hormone challenge versus controls. Additionally, the cortisol response tended to be greater overall (P < 0.10) in 15 versus 10-week-old pigs in response to the dexamethasone suppression test. The cortisol response tended to be reduced overall (P < 0.10) in 15 versus 10-week-old pigs in response to the corticotrophin releasing hormone challenge. Hypothalamic corticotropin releasing hormone mRNA abundance tended to be greater (P < 0.10) in in utero heat-stressed versus control pigs at 15-weeks of age. In summary, in utero heat stress altered some aspects of the hypothalamic-pituitary-adrenal axis related to corticotropin releasing hormone signaling, and age influenced this response.

Behavioral Response of Weaned Pigs during Gas Euthanasia with CO2, CO2 with Butorphanol, or Nitrous Oxide.

The swine industry is often forced to euthanize pigs in the first few weeks of life due to injuries, hernias, or unthriftiness. The majority of pigs are euthanized using carbon dioxide (CO2) gas asphyxiation but concerns as to the humaneness of CO2 are increasing. This study compared the euthanasia of weaned pigs using N2O (N2O; n = 9) or CO2 (n = 9), at 50% and 25% min-1 exchange rate, respectively. In addition, we administered an analgesic prior to euthanasia with CO2 (CO2B) exposure as a third treatment (n = 9) to elucidate behaviors indicative of pain. Pigs in the CO2 and N2O treatments lost posture at similar times (latency of 145.0 ± 17.3 and 162.6 ± 7.0 s respectively, p > 0.10), while the CO2B treatment pigs lost posture the soonest (101.2 ± 4.7 s, p < 0.01). The pigs in the CO2B treatment made more escape attempts than the CO2 or N2O pigs (16.4 ± 4.2, 4.7 ± 1.6, 0.3 ± 0.2, respectively; p < 0.0004). However, pigs in N2O squealed more often than either the CO2 or CO2B pigs (9.0 ± 1.6, 2.8 ± 1.2, 1.3 ± 0.6, respectively, p < 0.001). Given the similar time to loss of posture and shorter time displaying open mouth breathing, N2O may cause less stress to pigs; however, the greater number of squeals performed by these pigs suggests the opposite. It was not apparent that any behavior measured was indicative of pain. In conclusion, N2O applied at a 50% min-1 flow rate can be an alternative to CO2 for pig euthanasia.

Wireless 'under the skull' epidural EEG and behavior in piglets during nitrous oxide or carbon dioxide gas euthanasia.

Consciousness is central to animal welfare concerns. Its assessment is most often conducted based on behavior, with a poor understanding of the correspondence between behavior and the neurobiological processes that underlie the subjective experience of consciousness. Recording of brain electrical activity using electrodes placed under the skull improves EEG recording by minimizing artifacts from muscular or cardiac activities, and it can now be combined with wireless recording in free-moving animals. This experiment investigated the correspondence between wireless 'under the skull' epidural EEG and the behavior of 18 five-week-old female piglets undergoing nitrous oxide (N2O) or carbon dioxide (CO2) gradual fill gas euthanasia at 25% replacement rate per minute of the chamber volume. Piglets exposed to CO2 had a peak in EEG total power ('Ptot') during the flailing stage, whereas piglets exposed to N2O had a higher EEG 95% spectral edge frequency ('F95') during their initial explorative behavior phase and a drop in EEG median frequency ('F50') after loss of posture. Loss of posture without righting attempt, as the last behavioral state observed during euthanasia, preceded the onset of transitional EEG on average by 0.9 and 3.1 min (for CO2 and N2O treatments, respectively), and the onset of isoelectric EEG by 4.5 and 6.2 min (for CO2 and N2O treatments, respectively). Paddling movements occurred shortly before and during transitional EEG but never during isoelectric EEG, whereas gasps persisted after the EEG had become isoelectric. The dynamics of EEG spectral changes were complex to interpret in relation to the degree of consciousness, but isoelectric EEG as an unequivocal indicator of unconsciousness appeared several minutes after loss of posture with no righting attempt. This leaves a window of uncertainty in regards to the potential for consciousness after loss of posture during gradual fill gas euthanasia in piglets.

Evaluation and mitigation of the effects of in utero heat stress on piglet growth performance, postabsorptive metabolism, and stress response following weaning and transport.

In utero heat stress (IUHS) increases the energy requirements of pigs during postnatal life, and this may compound weaning and transport stress. The study objective was to evaluate and mitigate the negative effects of IUHS following weaning and transport through the provision of a nutrient-dense (ND) nursery diet formulated to meet the greater energy requirements of IUHS pigs during the first 14 d postweaning and transport. Twenty-four pregnant gilts were exposed to thermoneutral (TN; n = 12; 17.5 ± 2.1 °C) or heat stress (HS; n = 12; cycling 26 to 36 °C) conditions for the first half of gestation (day 6 to 59) and then TN conditions (20.9 ± 2.3 °C) until farrowing. Nine TN gilts and 12 HS gilts produced litters. At weaning (16.2 ± 0.4 d), mixed-sex piglets (N = 160; 4.78 ± 0.15 kg body weight [BW]) were transported (loading + transport + unloading) for 11 h 40 min. Following transport, piglets were blocked into pens (n = 4 pigs/pen) by in utero and dietary treatments: in utero thermoneutral (IUTN) + control (C) diet (n = 10 pens), IUTN + ND (n = 10 pens), IUHS + C (n = 10 pens), and IUHS + ND (n = 10 pens). Treatment diets were fed from day 1 to 14 postweaning and transport (period 1), and the C diet was fed to all pigs from day 14 to 35 postweaning and transport (period 2). Production measures were taken in 7 d intervals to calculate average daily gain (ADG), average daily feed intake (ADFI), average daily net energy intake (ADEI), gain:feed, and gain:net energy intake. Blood samples were collected prior to transport, following transport, and on days 2, 7, 14, 28, and 35 postweaning and transport to analyze cortisol, glucose, insulin, and nonesterified fatty acids. Behavior was assessed through video-recording on days 3, 5, 8, 11, and 13 postweaning and transport. In period 1, ADG was reduced (P = 0.04; 20.0 g/d) in IUHS vs. IUTN pigs. Pigs fed ND diets had reduced ADFI (P = 0.02; 9.3%) compared with C diet-fed pigs during period 1, which resulted in similar ADEI (P = 0.23; 1,115 ± 35 kcal/d). During transport, cortisol was decreased (P = 0.03; 25.8%) in IUHS vs. IUTN pigs. On day 2, glucose was decreased (P = 0.01; 13.8%) in IUHS vs. IUTN pigs. No in utero treatment-related behavior differences were observed but lying was reduced (P = 0.03; 6.5%) and standing was increased (P = 0.04; 14.1%) in ND vs. C pigs overall. In summary, IUHS reduced growth performance in pigs following weaning and transport, and providing an ND diet did not rescue the lost performance.

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-1ß (IL-1ß) 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-1ß 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.

Replacing dietary antibiotics with 0.20% l-glutamine in swine nursery diets: impact on health and productivity of pigs following weaning and transport1,2,3.

Antibiotic use has been limited in U.S. swine production. Therefore, the objective was to determine whether supplementing l-glutamine at cost-effective levels can replace dietary antibiotics to improve piglet welfare and productivity following weaning and transport. Based on previous research, we hypothesized that withholding dietary antibiotics would negatively affect pigs while diet supplementation with 0.20% l-glutamine (GLN) would have similar effects on pig performance and health as antibiotics. Mixed sex piglets (N = 480; 5.62 ± 0.06 kg BW) were weaned (18.4 ± 0.2 d of age) and transported for 12 h in central Indiana, for 2 replicates, during the summer of 2016 and the spring of 2017. Pigs were blocked by BW and allotted to 1 of 3 dietary treatments (n = 10 pens/dietary treatment/replicate [8 pigs/pen]); antibiotics (A; chlortetracycline [441 ppm] + tiamulin [38.6 ppm]), no antibiotics (NA), or GLN fed for 14 d. On days 15 to 34, pigs were provided common antibiotic-free diets in 2 phases. Data were analyzed using PROC MIXED in SAS 9.4. Day 14 BW and days 0 to 14 ADG were greater (P = 0.01) for A (5.6% and 18.5%, respectively) and GLN pigs (3.8% and 11.4%, respectively) compared with NA pigs, with no differences between A and GLN pigs. Days 0 to 14 ADFI increased for A (P < 0.04; 9.3%) compared with NA pigs; however, no differences were detected when comparing GLN with A and NA pigs. Once dietary treatments ceased, no differences (P > 0.05) in productivity between dietary treatments were detected. On day 13, plasma tumor necrosis factor alpha (TNF-α) was reduced (P = 0.02) in A (36.7 ± 6.9 pg/mL) and GLN pigs (40.9 ± 6.9 pg/mL) vs. NA pigs (63.2 ± 6.9 pg/mL). Aggressive behavior tended to be reduced overall (P = 0.09; 26.4%) in GLN compared with A pigs, but no differences were observed between A and GLN vs. NA pigs. Huddling, active, and eating/drinking behaviors were increased overall (P < 0.02; 179%, 37%, and 29%, respectively) in the spring replicate compared with the summer replicate. When hot carcass weight (HCW) was used as a covariate, loin depth and lean percentage were increased (P = 0.01; 4.0% and 1.1%, respectively) during the spring replicate compared with the summer replicate. In conclusion, GLN supplementation improved pig performance and health after weaning and transport similarly to A across replicates; however, the positive effects of A and GLN were diminished when dietary treatments ceased.

The impact of farrowing room noise on sows' reactivity to piglets.

Despite much interest in sow welfare, the impact of the acoustic environment on sow reactivity to her piglets is rarely considered. The objective of this study was to understand the impact of noise produced by mechanical ventilation and other sows on a sow's reactivity to her piglets. Sows were farrowed in one of three environments: 1) with eight other sows exposed to constant fan noise (GROUP-FAN; n = 10), 2) alone with fan noise present (ISO-FAN; n = 10), and 3) alone without fans running (ISO-QUIET; n = 10). Sows were subjected for 5 min to a piglet removal event (REMOVAL) by an unknown handler twice, at 24 and 48 h postfarrowing. During a REMOVAL, sows were observed via video recording for changes in posture, eating and drinking behavior, and head orientation. Audio was recorded to quantify vocalizations by the sow. Once piglets were returned, sows underwent further behavior observations for 10 min (RETURN), resulting in approximately 15 min of total video observation. Sows were classified as young (second and third parity) and old (fifth parity and older). The YOUNG sows tended to be more Alert (looking toward the handler or their piglets) during REMOVE than OLD sows (P = 0.07; 2.01 and 1.33, respectively). The ISO-FAN sows vocalized the loudest during REMOVAL (P < 0.001) with ISO-QUIET sows performing the quietest vocalizations (GROUP-FAN: 72.22 ± 1.06 dB; ISO-FAN: 73.61 ± 1.07 dB; ISO-QUIET: 67.41 ± 0.99 dB). During RETURN, YOUNG sows spent more time sitting than OLD sows (P < 0.01; 7.48 ± 1.6% and 0.91 ± 1.8%, respectively). The ISO-QUIET sows tended to have more posture changes during the RETURN with ISO-FAN having the least changes (P = 0.06; GROUP-FAN: 1.23 ± 0.4; ISO-FAN: 0.44 ± 0.3; ISO-QUIET: 1.61 ± 0.4). Finally, sows decreased the amount of time Alert in the second RETURN (P = 0.03; first: 3.9 ± 0.6%; second: 2.5 ± 0.6%). Overall, sows acclimated to the removal and return events with decreased vocalizations and decreased Alert behaviors in the second REMOVAL and RETURN. Additionally, YOUNG sows performed more active behaviors than OLD sows, indicating that sows may become less interested in or cannot hear their piglets as they age. Finally, there is some indication that ventilation presence has an effect on sow-piglet communication with ISO-FAN sows having the loudest vocalizations when compared with sows without ventilation noise, indicating that ventilation noise may be a possible competitor with a sow's ability to communicate with her piglets.

Auditory brainstem responses in weaning pigs and three ages of sows.

Piglet crushing is a devastating welfare concern on swine farms; however, some sows appear unresponsive to a piglet's call. Sow hearing ability is rarely considered despite the extensive body of research performed on crushing. In this study, pigs of four age groups (weaning, n = 7; gilts, n = 5; 2nd and 3rd parity, n = 5; 5th parity and up, n = 5) were anesthetized and auditory brainstem responses (ABRs) were performed to measure if pig hearing diminishes with age in a mechanically ventilated barn. Before testing, pigs were placed in a sound dampening box. ABRs were performed on animals using 1,000 clicks at two decibel (dB) levels: 90 and 127 dB sound pressure level. Latencies and amplitudes of waves I-V were measured and interpeak latencies for waves I-III, III-V, and I-V were calculated. Five pigs (three 2nd and 3rd parity, and two 5th parity and above) had no detectable waves at either decibel. Sows in 2nd and 3rd parities had very few distinguishable waves, with only wave I and II present in two sows. Amplitudes of waves I and V increased with increased dB (P < 0.001). Increasing dB decreased the latency of each of the recorded waves (P < 0.01). The vast majority of commercial swine are raised in noisy barn environments; it is possible that these environments directly affect the ability for pigs to hear and normal hearing development in this population of animals. Hearing has a significant effect on swine welfare as hearing is integral to successful animal handing and during moments of animal-to-animal communication. Hearing is a considerable welfare issue on farms and ways to decrease pig hearing loss should be considered.

Iron regulated genes of Salmonella enterica serovar Typhimurium in response to norepinephrine and the requirement of fepDGC for norepinephrine-enhanced growth.

Catecholamines may stimulate enteric bacteria including the foodborne pathogen Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) by two mechanisms in vivo: as a quorum sensing signal and a supplier of iron. To identify genes of Salmonella Typhimurium that respond to norepinephrine, transposon mutagenesis and DNA microarray analysis were performed. Insertional mutations in the following genes decreased norepinephrine-enhanced growth: degS, entE, entF, fes, gpmA, hfq, STM3846. DNA microarray and real-time RT-PCR analyses revealed a decrease in the expression of several genes involved in iron acquisition and utilization during norepinephrine exposure, signifying the iron-limiting conditions of serum-SAPI minimal medium and the siderophore-like activity of norepinephrine. Unlike the wild-type parent strain, growth of neither a fepA iroN cirA mutant nor a fepC mutant, harboring deletional mutations in the outer and inner membrane transporters of enterochelin, respectively, was enhanced by norepinephrine. However, growth of the fepC and the fepA iroN cirA mutants could be rescued by an alternative siderophore, ferrioxamine E, further validating the role of norepinephrine in supplying the organism with iron via the catecholate-specific iron transport system. Contrary to previous reports using small animal models, the fepA iroN cirA mutant of Salmonella Typhimurium colonized the swine gastrointestinal tract, as did the fepC mutant.

A Two-Step Process of Nitrous Oxide before Carbon Dioxide for Humanely Euthanizing Piglets: On-Farm Trials.

Current methods of euthanizing piglets are raising animal welfare concerns. Our experiment used a novel two-step euthanasia method, using nitrous oxide (N2O) for six minutes and then carbon dioxide (CO2) on compromised 0- to 7-day-old piglets. A commercial euthanasia chamber was modified to deliver two euthanasia treatments: the two-step method using N2O then CO2 (N2O treatment) or only CO2 (CO2 treatment). In Experiment 1, 18 piglets were individually euthanized. In Experiment 2, 18 groups of four to six piglets were euthanized. In the N2O treatment, piglets lost posture, indicating the onset of losing consciousness, before going into CO2 where they showed heavy breathing and open-mouth breathing; whereas piglets in the CO2 treatment did not lose posture until after exhibiting these behaviors (p ≤ 0.004). However, piglets in the N2O treatment took longer to lose posture compared to the CO2 treatment (p < 0.001). Piglets in the N2O treatment displayed more behavioral signs of stress and aversion: squeals/minute (p = 0.004), escape attempts per pig (p = 0.021), and righting responses per pig (p = 0.084) in a group setting. In these regards, it cannot be concluded that euthanizing piglets for 6 min with N2O and then CO2 is more humane than euthanizing with CO2 alone.

Comparison of Vocalization Patterns in Piglets Which Were Crushed to Those Which Underwent Human Restraint.

Though many studies focused on piglet crushing utilizing piglet vocalizations to test sow response, none have verified the properties of test vocalizations against actual crushing events. Ten sows were observed 48 h after parturition, and crushing events were recorded from all sows. When a crushing event occurred, a second piglet within the same litter was used to solicit a vocalization through manual restraint to compare restrained piglets' call properties to those of crushed piglets'. A total of 659 Restrained calls and 631 Crushed calls were collected. Variables were gathered at the loudest point in a call, and as an average across the entire call. Crushed piglets had a lower fundamental frequency (p < 0.01; Crushed: 523.57 ± 210.6 Hz; Restrained: 1214.86 ± 203.2 Hz) and narrower bandwidth (p < 0.01; Crushed: 4897.01 ± 587.3 Hz; Restrained: 6674.99 ± 574.0 Hz) when analyzed at the loudest portion of a call. Overall, piglets which were crushed had a lower mean peak frequency than those which were restrained (p = 0.01; 1497.08 ± 239.4 Hz and 2566.12 ± 235.0 Hz, respectively). Future research should focus on measuring sow reactivity to Crushed and Restrained piglets to continue to improve research practices.