Impact of Arachidonic and Docosahexaenoic Acid Supplementation on Neural and Immune Development in the Young Pig.

Background: Human milk contains both arachidonic acid (ARA) and docosahexaenoic acid (DHA). Supplementation of infant formula with ARA and DHA results in fatty acid (FA) profiles, neurodevelopmental outcomes, and immune responses in formula-fed infants that are more like those observed in breastfed infants. Consequently, ARA and DHA have been historically added together to infant formula. This study investigated the impact of ARA or DHA supplementation alone or in combination on tissue FA incorporation, immune responses, and neurodevelopment in the young pig. Methods: Male pigs (N = 48 total) received one of four dietary treatments from postnatal day (PND) 2-30. Treatments targeted the following ARA/DHA levels (% of total FA): CON (0.00/0.00), ARA (0.80/0.00), DHA (0.00/0.80), and ARA+DHA (0.80/0.80). Plasma, red blood cells (RBC), and prefrontal cortex (PFC) were collected for FA analysis. Blood was collected for T cell immunophenotyping and to quantify a panel of immune outcomes. Myelin thickness in the corpus callosum was measured by transmission electron microscopy and pig movement was measured by actigraphy. Results: There were no differences in formula intake or growth between dietary groups. DHA supplementation increased brain DHA, but decreased ARA, compared with all other groups. ARA supplementation increased brain ARA compared with all other groups but did not affect brain DHA. Combined supplementation increased brain DHA levels but did not affect brain ARA levels compared with the control. Pigs fed ARA or ARA+DHA exhibited more activity than those fed CON or DHA. Diet-dependent differences in activity suggested pigs fed ARA had the lowest percent time asleep, while those fed DHA had the highest. No differences were observed for immune or myelination outcomes. Conclusion: Supplementation with ARA and DHA did not differentially affect immune responses, but ARA levels in RBC and PFC were reduced when DHA was provided without ARA. Supplementation of either ARA or DHA alone induced differences in time spent asleep, and ARA inclusion increased general activity. Therefore, the current data support the combined supplementation with both ARA and DHA in infant formula and raise questions regarding the safety and nutritional suitability of ARA or DHA supplementation individually.

Dietary osteopontin-enriched algal protein as nutritional support in weaned pigs infected with F18-fimbriated enterotoxigenic Escherichia coli.

This study investigated the effects of dietary osteopontin (OPN)-enriched algal protein on growth, immune status, and fecal fermentation profiles of weaned pigs challenged with a live infection of F18-fimbriated enterotoxigenic E. coli (ETEC). At 21 d of age, 54 pigs (5.95 ± 0.28 kg BW; blocked by BW) were allotted to 1 of 3 experimental groups combining dietary and health statuses. A control diet, containing 1% wild-type algal protein, was fed to both sham-inoculated (NC) and ETEC-inoculated (PC) pigs, while the test diet contained 1% OPN-enriched algal protein as fed only to ETEC-inoculated pigs (OA). All pigs received their assigned dietary treatment starting at study initiation to permit a 10-d acclimation period prior to inoculation. Growth performance, fecal dry matter, as well as hematological, histopathological, immune, and microbiota outcomes were analyzed by ANOVA, where treatment and time were considered as fixed effects and pig as a random effect; significance was accepted at P < 0.05. Overall, ETEC-inoculated pigs (PC and OA) exhibited decreased (P < 0.05) ADG and G:F, as well as increased (P < 0.05) peripheral blood helper T-cells and total leukocyte counts, compared with NC pigs during the postinoculation period. The OA treatment also elicited the highest (P < 0.05) concentrations of circulating tumor necrosis factor-α and volatile fatty acid concentrations in luminal contents at various postinoculation time-points, compared with other treatments. A principal coordinate analysis based on Unifrac weighted distances indicated that NC and OA groups had similar overall bacterial community structures, while PC pigs exhibited greater diversity, but infection status had no impact on α-diversity. Osteopontin-specific effects on microbial community structure included enrichment within Streptococcus and Blautia genera and decreased abundance of 12 other genera as compared with PC pigs. Overall, ETEC-infected pigs receiving 1% OPN-enriched algal protein exhibited changes immunity, inflammatory status, and colonic microbial community structure that may benefit weanling pigs experiencing F18 ETEC infection.

Alterations of fecal microbiome characteristics by dietary soy isoflavone ingestion in growing pigs infected with porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important disease, and the ingestion of soy isoflavones (ISF) may benefit PRRSV-infected pigs due to demonstrated anti-inflammatory and antiviral properties. The objective of this study was to quantify the effects of ISF consumption on fecal microbiome characteristics at different timepoints across a disease challenge and determine whether any changes, if present, elude to potential biological mechanisms for previously observed performance benefits. In total, 96 weaned barrows were group-housed in a Biosafety Level-2 containment facility and allotted to one of three experimental treatments that were maintained throughout the study: noninfected pigs receiving an ISF-devoid control diet (NEG, n = 24) and infected pigs receiving either the control diet (POS, n = 36) or that supplemented with total ISF in excess of 1,600 mg/kg (ISF, n = 36). Following a 7-d adaptation, pigs were inoculated intranasally with either a sham-control (phosphate-buffered saline) or live PRRSV (1 × 105 median tissue culture infectious dose[TCID]50/mL, strain NADC20). Fecal samples were collected from 48 individual pigs at pre-infection (-2 d post-inoculation [DPI]), peak-infection (10 DPI), and post-infection (144 DPI) timepoints. Extracted DNA was used to quantify fecal microbiota profiles via 16S bacterial rRNA sequencing. Differences in bacterial communities among diet groups were evaluated with principal coordinate analysis and permutational multivariate analysis of variance using UniFrac distance matrices based on both unweighted and weighted UniFrac distances using QIIME 2. All other data were analyzed by one-way ANOVA performed on square root transformations using R. Across all timepoints, only a few differences were observed due to ISF alone mainly in lowly abundant genera. The most notable differences observed were decreased relative abundance of Actinobacteria at 144 DPI between noninfected and infected treatments (P < 0.05), which is consistent with various dysbioses observed in other disease models. Our findings indicate that the differences present were mainly due to PRRSV-infection alone and not strongly influenced by diet, which implies that previously observed performance benefits conferred by dietary ISF are not likely due to the changes in microbiome composition.

Effect of porcine reproductive and respiratory syndrome virus infection and soy isoflavone supplementation on carcass cutability and meat quality of pigs.

The objective was to evaluate the effects of porcine reproductive and respiratory syndrome virus (PRRSV) infection and dietary soy isoflavone (ISF) supplementation on carcass cutability and meat quality of commercial pigs. Barrows (21 d of age) were randomly allotted to experimental treatments that were maintained throughout the study: noninfected pigs received an ISF-devoid control diet (CON, n = 22) and infected pigs received either the control diet (PRRSV-CON, n = 20) or that supplemented with total ISF in excess of 1,500 mg/kg (PRRSV-ISF, n = 25). Pigs were penned by treatment, with six pigs within a pen. Following a 7-d adaptation, weanling pigs were inoculated once intranasally with either a sham-control (phosphate buffered saline [PBS]) or live PRRSV (1 × 105 tissue culture infective dose [TCID]50/mL, strain NADC20). Pigs were maintained on experimental diets for 166 d after inoculation and then slaughtered (192 or 194 d of age; approximately 120 kg body weight [BW]). At 1-d postmortem, left sides were separated between the 10th and 11th rib for the determination of loin eye area (LEA), backfat (BF) thickness, and loin quality (ultimate pH, instrumental color, drip loss, visual color, marbling, and firmness). Loin chops were aged 14 d postmortem prior to Warner-Bratzler shear force (WBSF) determination. Belly width, length, thickness, and flop distance were determined. Data were analyzed as a one-way ANOVA with pig as the experimental unit. Carcass yield, LEA, BF, and estimated lean percentage did not differ (P > 0.26) among treatments. Loins from CON pigs had increased ultimate pH (P = 0.01), reduced L* scores (P = 0.005) coupled with darker visual color scores (P = 0.004), were firmer (P < 0.0001), and exhibited reduced drip loss (P = 0.01) compared with PRRSV-CON and PRRSV-ISF pigs. However, WBSF did not differ (P = 0.51) among treatments after 14 d of aging. Bellies from CON pigs were more firm compared with bellies from PRRSV-CON and ISF pigs (P < 0.01). These data suggest PRRSV infection did not alter carcass characteristics but may have marginally reduced loin and belly quality. Supplementation with dietary soy isoflavones did nothing to mitigate the detrimental effects of PRRSV infection.

Dietary soy isoflavones reduce pathogen-related mortality in growing pigs under porcine reproductive and respiratory syndrome viral challenge.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important disease, and ingestion of soy isoflavones (ISF) may benefit PRRSV-infected pigs due to demonstrated anti-inflammatory and antiviral properties. The objective of this experiment was to recreate immunological effects previously observed in young pigs infected with PRRSV receiving ISF and determine how those effects influence growth performance during the entire growth period from weaning to market. In total, 96 weaned barrows were group housed in a biosafety level-2 containment facility and allotted to 1 of 3 experimental treatments that were maintained throughout the study: noninfected pigs received an ISF-devoid control diet (NEG, n = 24), and infected pigs received either the control diet (POS, n = 36) or that supplemented with total ISF in excess of 1,600 mg/kg (ISF, n = 36). Following a 7-d adaptation, weanling pigs were inoculated intranasally with either a sham-control (PBS) or live PRRSV (1 × 105 TCID50/mL, strain NADC20). After inoculation, individual blood samples (n = 8 to 12/treatment) were routinely collected to monitor viral clearance and hematological parameters, including serum neutralizing anti-PRRSV antibody production. Pen-based oral fluids were used to monitor PRRSV clearance at later growth stages. A 1- or 2-way ANOVA was performed to compare experimental treatments depending on whether the outcome was repeatedly measured. In general, PRRSV infection decreased performance during early growth phases, resulting in 5.4% lower final BW for POS vs. NEG pigs (P < 0.05). Dietary ISF elicited inconsistent effects on growth performance, increased (P < 0.05) neutrophil cell counts and the relative proportion of memory T-cells, and decreased (P < 0.05) the time to full PRRSV clearance from oral fluids. Dietary ISF also elicited earlier, more robust anti-PRRSV neutralizing antibody production when compared with POS pigs. Additionally, and most notably, POS pigs experienced ~50% greater infection-related mortality rate vs. ISF pigs (P < 0.05), which may have significant economic implications for producers. Overall, dietary ISF ingestion supported immune responses and reduced mortality in PRRSV-infected pigs when fed to growing pigs though the biological mechanism of these effects remains unclear.

TECHNICAL NOTE: A method for detection of differences in cook loss and tenderness of aged pork chops cooked to differing degrees of doneness using sous-vide.

The objective was to determine the ability to detect differences in cook loss and Warner-Bratzler shear force (WBSF) values between chops aged for differing time periods and cooked to varying degrees of doneness with in a sous-vide style cooker. Loins from pigs (HCW = 96 kg) humanely slaughtered at the University of Illinois Meat Science Laboratory were separated between the 10th and 11th rib into anterior and posterior sections. The posterior section was cut into 6 separate 2.54-cm-thick chops. The middle 4 chops were randomly designated for aging of 3 d and cooked to 63 °C, aged 7 d and cooked to 63 °C, aged 14 d and cooked to 63 °C, or aged 14 d and cooked to 71 °C. Chops were cooked by placing them in a water bath with an immersion circulator set to the desired end-point temperature for 90 min. Cook loss was calculated for each chop by measuring initial and final weight, and accounting for packaging weight. Four cores measuring 1.25 cm in diameter were cut parallel to the muscle fibers from each chop and analyzed for WBSF. Data were analyzed using a 1-way ANOVA. Least squares means were separated using the probability of difference option in the MIXED procedure of SAS. Among chops cooked to 63 °C, chops aged 3 d has less (P < 0.01) cook loss than those aged 7 d, and chops aged 7 d had less (P < 0.01) cook loss than those aged 14 d. Among chops aged for 14 d, chops cooked to 71 °C had greater (P < 0.001) cook loss than chops cooked to 63 °C. Differences in tenderness were also detected between aging periods. Among chops cooked to 63 °C, chops aged 3 d required more (P = 0.02) force to shear than those aged 7 d, but chops aged 7 d did not differ (P = 0.15) from those aged 14 d. Chops aged 14 d and cooked to 71 °C required (P < 0.0001) more force than those aged 14 d and cooked to 63 °C. Overall, these data indicate that sous-vide is an acceptable cooking method for use in experiments as expected differences in cook loss and WBSF were detected in chops aged to differing time points or cooked to differed degrees of doneness.

The role of the cytoskeleton in cellular force generation in 2D and 3D environments.

To adhere and migrate, cells generate forces through the cytoskeleton that are transmitted to the surrounding matrix. While cellular force generation has been studied on 2D substrates, less is known about cytoskeletal-mediated traction forces of cells embedded in more in vivo-like 3D matrices. Recent studies have revealed important differences between the cytoskeletal structure, adhesion, and migration of cells in 2D and 3D. Because the cytoskeleton mediates force, we sought to directly compare the role of the cytoskeleton in modulating cell force in 2D and 3D. MDA-MB-231 cells were treated with agents that perturbed actin, microtubules, or myosin, and analyzed for changes in cytoskeletal organization and force generation in both 2D and 3D. To quantify traction stresses in 2D, traction force microscopy was used; in 3D, force was assessed based on single cell-mediated collagen fibril reorganization imaged using confocal reflectance microscopy. Interestingly, even though previous studies have observed differences in cell behaviors like migration in 2D and 3D, our data indicate that forces generated on 2D substrates correlate with forces within 3D matrices. Disruption of actin, myosin or microtubules in either 2D or 3D microenvironments disrupts cell-generated force. These data suggest that despite differences in cytoskeletal organization in 2D and 3D, actin, microtubules and myosin contribute to contractility and matrix reorganization similarly in both microenvironments.