Porcine epidemic diarrhea virus infection induces endoplasmic reticulum stress and unfolded protein response in jejunal epithelial cells of weaned pigs.

Porcine epidemic diarrhea virus (PEDV) infection leads to diarrhea and subsequently to decreased feed efficiency and growth in weaned pigs. Given that few studies have addressed the host-virus interaction in vivo, this study focused on endoplasmic reticulum (ER) stress and unfolded protein response (UPR) in jejunal epithelial cells during PEDV infection. Eight-week-old pigs (n = 64) were orally inoculated with PEDV IN19338 strain (n = 40) or sham-inoculated (n = 24) and analyzed for PEDV viral RNA shedding using reverse transcription-quantitative polymerase chain reaction and for viral antigen within enterocytes using immunohistochemistry (IHC). ER stress was analyzed in a subset of 9 PEDV-inoculated pigs with diarrhea, detectable viral RNA, and viral antigen (PEDV-immunopositive pigs). Compared with control pigs, PEDV-immunopositive pigs had a reduced ratio of villus height to crypt depth in the jejunum (P = .002, n = 9 per group), consistent with intestinal injury. The protein levels of ATF6, IRE1, PERK, XBP1u, ATF4, GRP78, and caspase-3 were assessed in jejunal epithelial cells at the villus tips via IHC. Both ER stress and UPR were demonstrated in PEDV-immunopositive pigs by the increased expression of ATF6 (P = .047), IRE1 (P = .007), and ATF4 (P = .001). The expression of GRP78 (P = .024) and caspase-3 (P = .004) were also increased, indicating an accompanying increase in ER protein folding capacity and apoptosis. Overall, these results reveal that PEDV infection induces ER stress and UPR in intestinal epithelial cells of weaned pigs.

BONE MINERAL DENSITY DETERMINED BY DUAL-ENERGY X-RAY ABSORPTIOMETRY METHOD FOR EGYPTIAN FRUIT BATS (ROUSETTUS AEGYPTIACUS) IN HUMAN CARE.

Egyptian fruit bats (Rousettus aegyptiacus) are commonly held within zoos and research facilities. Despite this popularity, there have been no publications regarding normal bone density or bone mineral concentration and few publications regarding the normal dietary requirements of calcium, phosphorus, and vitamin D (25-hydrox-vitamin D3 [25(OH)D3]) for the species. A clinical investigation into frequent fractures in a zoo population of 23 adult male Egyptian fruit bats used dual-energy X-ray absorptiometry (DXA) to evaluate bone mineral density (BMD) and bone mineral content (BMC) prior to and after dietary adjustment of calcium and phosphorus. This study documents BMD, BMC, serum trace minerals, and serum levels of 25(OH)D3 within this population and includes postmortem bone density and bone ash from a subset of bats that died during the study period. A handheld point-of-care analyzer was utilized to assess ionized calcium and electrolytes (sodium, potassium, chloride, total CO2), glucose, blood urea nitrogen, and creatinine at each DXA time point to contribute to published reference ranges for this species. This study identified a significant (P < 0.001) increase in BMC over 15 mon (4.71-8.26 g) following additional calcium and phosphorus supplementation in the diet. The BMD also trended toward a significant increase (0.124-0.274 g/cm2, P < 0.053). Additional studies documenting normal bone density that include both sexes and possibly free-ranging bats consuming a natural diet are recommended to determine normal values for R. aegyptiacus.

Lawsonia intracellularis infected enterocytes lack sucrase-isomaltase which contributes to reduced pig digestive capacity.

Lawsonia intracellularis is endemic to swine herds worldwide, however much is still unknown regarding its impact on intestinal function. Thus, this study aimed to characterize the impact of L. intracellularis on digestive function, and how vaccination mitigates these impacts. Thirty-six L. intracellularis negative barrows were assigned to treatment groups (n = 12/trt): (1) nonvaccinated, L. intracellularis negative (NC); (2) nonvaccinated, L intracellularis challenged (PC); and (3) L. intracellularis challenged, vaccinated (Enterisol® Ileitis, Boehringer Ingelheim) 7 weeks pre-challenge (VAC). On days post-inoculation (dpi) 0 PC and VAC pigs were inoculated with L. intracellularis. From dpi 19-21 fecal samples were collected for apparent total tract digestibility (ATTD) and at dpi 21, pigs were euthanized for sample collection. Post-inoculation, ADG was reduced in PC pigs compared with NC (41%, P < 0.001) and VAC (25%, P < 0.001) pigs. Ileal gross lesion severity was greater in PC pigs compared with NC (P = 0.003) and VAC (P = 0.018) pigs. Dry matter, organic matter, nitrogen, and energy ATTD were reduced in PC pigs compared with NC pigs (P ≤ 0.001 for all). RNAscope in situ hybridization revealed abolition of sucrase-isomaltase transcript in the ileum of PC pigs compared with NC and VAC pigs (P < 0.01). Conversely, abundance of stem cell signaling markers Wnt3, Hes1, and p27Kip1 were increased in PC pigs compared with NC pigs (P ≤ 0.085). Taken together, these data demonstrate that reduced digestibility during L. intracellularis challenge is partially driven by abolition of digestive machinery in lesioned tissue. Further, vaccination mitigated several of these effects, likely from lower bacterial burden and reduced disease severity.

Acute systemic inflammatory response to lipopolysaccharide stimulation in pigs divergently selected for residual feed intake.

BACKGROUND: It is unclear whether improving feed efficiency by selection for low residual feed intake (RFI) compromises pigs' immunocompetence. Here, we aimed at investigating whether pig lines divergently selected for RFI had different inflammatory responses to lipopolysaccharide (LPS) exposure, regarding to clinical presentations and transcriptomic changes in peripheral blood cells. RESULTS: LPS injection induced acute systemic inflammation in both the low-RFI and high-RFI line (n = 8 per line). At 4 h post injection (hpi), the low-RFI line had a significantly lower (p = 0.0075) mean rectal temperature compared to the high-RFI line. However, no significant differences in complete blood count or levels of several plasma cytokines were detected between the two lines. Profiling blood transcriptomes at 0, 2, 6, and 24 hpi by RNA-sequencing revealed that LPS induced dramatic transcriptional changes, with 6296 genes differentially expressed at at least one time point post injection relative to baseline in at least one line (n = 4 per line) (|log2(fold change)| ≥ log2(1.2); q < 0.05). Furthermore, applying the same cutoffs, we detected 334 genes differentially expressed between the two lines at at least one time point, including 33 genes differentially expressed between the two lines at baseline. But no significant line-by-time interaction effects were detected. Genes involved in protein translation, defense response, immune response, and signaling were enriched in different co-expression clusters of genes responsive to LPS stimulation. The two lines were largely similar in their peripheral blood transcriptomic responses to LPS stimulation at the pathway level, although the low-RFI line had a slightly lower level of inflammatory response than the high-RFI line from 2 to 6 hpi and a slightly higher level of inflammatory response than the high-RFI line at 24 hpi. CONCLUSIONS: The pig lines divergently selected for RFI had a largely similar response to LPS stimulation. However, the low-RFI line had a relatively lower-level, but longer-lasting, inflammatory response compared to the high-RFI line. Our results suggest selection for feed efficient pigs does not significantly compromise a pig's acute systemic inflammatory response to LPS, although slight differences in intensity and duration may occur.

Short-term heat stress results in increased apoptotic signaling and autophagy in oxidative skeletal muscle in Sus scrofa.

Prolonged environment-induced hyperthermia causes morbidities and mortality in humans and animals and appears to cause organ-specific injury and dysfunction. We have previously determined autophagic dysfunction and apoptotic signaling in oxidative skeletal muscle following prolonged hyperthermia. The aim of this investigation was to extend our knowledge regarding the early chronology of heat stress-mediated apoptotic and autophagic signaling in oxidative skeletal muscle. We hypothesized that 2, 4, and 6 h of hyperthermia would increase apoptosis and autophagy in oxidative skeletal muscle compared to thermoneutral (TN) conditions. Pigs were assigned to four groups (n = 8/group) and exposed to environmental heat stress (37 °C) for 0, 2, 4, or 6 h. Immediately following environmental exposure animals were euthanized and the red portion of the semitendinosus was collected. Markers of apoptotic signaling were increased following 2 h of heating but returned to baseline thereafter, while caspase 3 activity remained elevated 2-3 fold (p < .05) throughout the hyperthermic period. Heat stress increased (p < .05) markers of autophagic activation, and nucleation as well as autophagosome formation and degradation linearly throughout the heating intervention. In addition, 6 h of hyperthermia increased (p < .05) markers of mitophagy. These data suggest that apoptotic signaling precedes increased autophagy during acute heat stress in oxidative skeletal muscle.

Twelve hours of heat stress induces inflammatory signaling in porcine skeletal muscle.

Heat stress causes morbidity and mortality in humans and animals and threatens food security by limiting livestock productivity. Inflammatory signaling may contribute to heat stress-mediated skeletal muscle dysfunction. Previously, we discovered increased circulating endotoxin and intramuscular oxidative stress and TNF-α protein abundance, but not inflammatory signaling following 24 and 72 h of heat stress. Thus the purpose of this investigation was to clarify the role of inflammatory signaling in heat-stressed skeletal muscle. Crossbred gilts (n = 8/group) were assigned to either thermal neutral (24°C), heat stress (37°C), or pair-fed thermal neutral (24°C) conditions for 12 h. Following treatment, animals were euthanized, and the semitendinosus red (STR) and white (STW) were recovered. Heat stress did not alter inflammatory signaling in STW. In STR, relative heat shock protein abundance was similar between groups, as was nuclear content of heat shock factor 1. In whole homogenate, relative abundance of the NF-κB activator inhibitory κB kinase-α was increased by heat stress, although abundance of NF-κB was similar between groups. Relative abundance of phosphorylated NF-κB was increased by heat stress in nuclear fractions. Activator protein-1 (AP-1) signaling was similar between groups. While there were few differences in transcript expression between thermal neutral and heat stress, 80 and 56% of measured transcripts driven by NF-κB or AP-1, respectively, were increased by heat stress compared with pair-fed thermal neutral. Heat stress also caused a reduction in IL-6 transcript and relative protein abundance. These data demonstrate that short-term heat stress causes inflammatory signaling through NF-κB in oxidative, but not glycolytic, skeletal muscle.

Selenium and vitamin E together improve intestinal epithelial barrier function and alleviate oxidative stress in heat-stressed pigs.

What is the central question of this study? Oxidative stress may play a role in compromising intestinal epithelial barrier integrity in pigs subjected to heat stress, but it is unknown whether an increase of dietary antioxidants (selenium and vitamin E) could alleviate gut leakiness in heat-stressed pigs. What is the main finding and its importance? Levels of dietary selenium (1.0 p.p.m.) and vitamin E (200 IU kg(-1) ) greater than those usually recommended for pigs reduced intestinal leakiness caused by heat stress. This finding suggests that oxidative stress plays a role in compromising intestinal epithelial barrier integrity in heat-stressed pigs and also provides a nutritional strategy for mitigating these effects. Heat stress compromises the intestinal epithelial barrier integrity of mammals through mechanisms that may include oxidative stress. Our objective was to test whether dietary supplementation with antioxidants, selenium (Se) and vitamin E (VE), protects intestinal epithelial barrier integrity in heat-stressed pigs. Female growing pigs (n = 48) were randomly assigned to four diets containing from 0.2 p.p.m. Se and 17 IU kg(-1) VE (control, National Research Council recommended) to 1.0 p.p.m. Se and 200 IU kg(-1) VE for 14 days. Six pigs from each dietary treatment were then exposed to either thermoneutral (20°C) or heat-stress conditions (35°C 09.00-17.00 h and 28°C overnight) for 2 days. Transepithelial electrical resistance and fluorescein isothiocyanate-dextran (4 kDa; FD4) permeability were measured in isolated jejunum and ileum using Ussing chambers. Rectal temperature, respiratory rate and intestinal HSP70 mRNA abundance increased (all P < 0.001), and respiratory alkalosis occurred, suggesting that pigs were heat stressed. Heat stress also increased FD4 permeability and decreased transepithelial electrical resistance (both P < 0.01). These changes were associated with changes indicative of oxidative stress, a decreased glutathione peroxidase (GPX) activity and an increased glutathione disulfide (GSSG)-to-glutathione (GSH) ratio (both P < 0.05). With increasing dosage of Se and VE, GPX-2 mRNA (P = 0.003) and GPX activity (P = 0.049) increased linearly, the GSSG:GSH ratio decreased linearly (P = 0.037), and the impacts of heat stress on intestinal barrier function were reduced (P < 0.05 for both transepithelial electrical resistance and FD4 permeability). In conclusion, in pigs an increase of dietary Se and VE mitigated the impacts of heat stress on intestinal barrier integrity, associated with a reduction in oxidative stress.

Postprandial serum endotoxin in healthy humans is modulated by dietary fat in a randomized, controlled, cross-over study.

BACKGROUND: High-fat diets may contribute to metabolic disease via postprandial changes in serum endotoxin and inflammation. It is unclear how dietary fat composition may alter these parameters. We hypothesized that a meal rich in n-3 (ω3) fatty acids would reduce endotoxemia and associated inflammation but a saturated or n-6 (ω6) fatty acid-rich meal would increase postprandial serum endotoxin concentrations and systemic inflammation in healthy adults. METHODS: Healthy adults (n = 20; mean age 25 ± 3.2 S.D. years) were enrolled in this single-blind, randomized, cross-over study. Participants were randomized to treatment and reported to the laboratory, after an overnight fast, on four occasions separated by at least one week. Participants were blinded to treatment meal and consumed one of four isoenergetic meals that provided: 1) 20 % fat (control; olive oil) or 35 % fat provided from 2) n-3 (ω3) (DHA = 500 mg; fish oil); 3) n-6 (ω6) (7.4 g; grapeseed oil) or 4) saturated fat (16 g; coconut oil). Baseline and postprandial blood samples were collected. Primary outcome was defined as the effect of treatment meal on postprandial endotoxemia. Serum was analyzed for metabolites, inflammatory markers, and endotoxin. Data from all 20 participants were analyzed using repeated-measures ANCOVA. RESULTS: Participant serum endotoxin concentration was increased during the postprandial period after the consumption of the saturated fat meal but decreased after the n-3 meal (p < 0.05). The n-6 meal did not effect a different outcome in participant postprandial serum endotoxin concentration from that of the control meal (p > 0.05). There was no treatment meal effect on participant postprandial serum biomarkers of inflammation. Postprandial serum triacylglycerols were significantly elevated following the n-6 meal compared to the n-3 meal. Non-esterified fatty acids were significantly increased after consumption of the saturated fat meal compared to other treatment meals. CONCLUSIONS: Meal fatty acid composition modulates postprandial serum endotoxin concentration in healthy adults. However, postprandial endotoxin was not associated with systemic inflammation in vivo. TRIAL REGISTRATION: This study was retrospectively registered at clinicaltrials.gov as NCT02521779 on July 28, 2015.

Effects of dietary protein level on intestinal function and inflammation in nursery pigs.

High crude protein (CP; 21% to 26%) diets fed during the first 21 to 28 d postweaning are viewed negatively because of a perceived increase in the incidence rates of diarrhea due to increased intestinal protein fermentation and/or augmented enteric pathogen burden. This is thought to antagonize nursery pig health and growth performance. Therefore, our objective was to evaluate the impact of low vs. high dietary CP on 21-day postweaned pig intestinal function. Analyzed parameters included ex vivo intestinal barrier integrity (ileum and colon), ileal nutrient transport, tissue inflammation, and fecal DM. One hundred and twenty gilts and barrows (average body weight) were randomly assigned to one of two diets postweaning. Diets were fed for 21 d, in two phases. Phase 1 diets: low CP (17%) with a 1.4% standardized ileal digestible (SID) Lys (LCP), or high CP (24%) with a 1.4% SID Lysine (HCP). Phase 2: LCP (17%) and a 1.35% SID lysine, or HCP (24%) formulated to a 1.35% SID lysine. Pig growth rates, feed intakes, and fecal consistency did not differ (P > 0.05) due to dietary treatment. Six animals per treatment were euthanized for additional analyses. There were no differences in colonic epithelial barrier function as measured by transepithelial electrical resistance (TER) and fluorescein isothiocyanate (FITC)-dextran transport between treatments (P > 0.05). Interleukins (IL)-1α, IL-1ß, IL-1ra, IL-2 IL-4, IL-6, and IL-12 were not different between treatments (P > 0.05). However, IL-8 and IL-18 were higher in HCP- vs. LCP-fed pigs (P < 0.05). There were no differences in fecal dry matter (DM; P > 0.05) between treatments. In the ileum, there was a tendency (P = 0.06) for TER to be higher in HCP-fed pigs, suggesting a more robust barrier. Interestingly, glucose and glutamine transport were decreased in HCP- vs. LCP-fed pigs (P < 0.05). FITC-dextran transport was not different between treatments (P > 0.05). There were also no differences in ileal cytokine concentrations between diets (P > 0.05). Taken together, the data show that low CP does not negatively impact colonic barrier function, fecal DM, or inflammation. In contrast, ileal barrier function and nutrient transport were altered, suggesting a regional effect of diet on overall intestinal function.

High dietary crude protein (CP) is thought to antagonize nursery pig enteric health. Feeding high CP diets to nursery pigs did not exacerbate intestinal health or inflammation, and overall, protein level in the diet has little impact on animal health and performance.

Soluble corn fiber, resistant corn starch, and protected butyrate effects on performance, gastrointestinal volatile fatty acids, and apparent total-tract digestibility of calcium and phosphorus in nursery pigs.

An experiment was conducted to determine how feeding calcium (Ca)-deficient diet would affect gastrointestinal pH and volatile fatty acids (VFAs), Ca digestibility, bone mineral density (BMD), and performance in nursery pigs; and if supplementation of nondigestible oligosaccharides would affect these same parameters. In total, 240 weaned pigs (BW = 7.1 kg) were placed into 80 pens with 3 pigs/pen. The eight dietary treatments consisted of: 1) positive control (PC, 0.83% total Ca), 2) negative control (NC, 0.50% total Ca), 3 and 4) NC + 5% or 7.5% soluble corn fiber (SCF), 5 and 6) NC + 5% or 7.5% resistant corn starch (rCS), 7 and 8) NC + 0.25% or 0.50% fat-protected butyrate (pBRT). Pigs were ad libitum fed the dietary treatments for 21 d to determine average daily gain (ADG), average daily feed intake (ADFI) and gain:feed ratio (GF) with a fecal sample collected from each pen to determine Ca digestibility using acid insoluble ash as the dietary marker, with 1 pig/pen euthanized on d 21 for collection of ileal and colon contents and the left humerus. Pigs fed the NC diet had a lower colonic pH compared with pigs fed the PC (P = 0.06) but no effect on total VFA was observed (P > 0.10). Pigs fed diets containing SCF and rCS had lower colonic pH and total VFA compared to pigs fed the NC diet (P ≤ 0.05). Pigs fed diets containing pBRT had greater colonic total VFA compared to pigs fed the NC diet (P ≤ 0.07), but no difference in colonic pH was observed (P > 0.10). Pigs fed the NC diet had a greater Ca digestibility compared to pigs fed the PC (P ≤ 0.01), with no treatment to the NC having any effect on Ca digestibility compared to pigs fed the NC (P > 0.10). There was no effect of dietary Ca level on BMD and no overall addition of feeding SCF, rCS, or pBRT on BMD compared to pigs fed the NC (P > 0.10). There was no impact on pig ADG, ADFI, or GF by reducing dietary Ca by 40% (i.e., pigs fed the NC) compared to pigs fed the PC (P > 0.10). Relative to pigs fed the NC, there was no overall effect of SCF, rCS, or pBRT on ADG, ADFI, or GF (P > 0.10). In conclusion, feeding young pigs a Ca-deficient diet reduced colonic pH, increased digestibility of Ca, but had no impact on bone mineralization or overall pig performance. Supplementation of nondigestible oligosaccharides pr protected butyrate had either no effect or an inconsistent effect on colonic pH, Ca, or PHOS digestibility, bone mineralization, or overall pig performance.

Calcium (Ca) is a major component of the skeleton in addition to being essential for growth and is imperative for bone mass development. Improvement in Ca absorption in Ca-deficient diets has been shown in human and rodent studies when nondigestible oligosaccharides have been consumed due to a modification of gastrointestinal conditions which increase mineral solubility. Because swine have been shown to be an excellent model for human nutrition research, an experiment was conducted to determine how a moderately Ca-deficient diet would affect gastrointestinal fermentation conditions, Ca and phosphorus (PHOS) digestibility, bone mineralization, and growth performance in nursery pigs; and if supplementation of nondigestible oligosaccharides would affect these same parameters. Results indicate that feeding young pigs a diet below recommended levels of Ca reduced colonic pH, increased apparent total-tract digestibility of Ca and PHOS, but had no impact on bone mineralization or overall pig performance. Supplementation of nondigestible oligosaccharides had inconsistent effects on colonic pH, and did not affect Ca or PHOS digestibility, bone mineralization, or overall pig performance.

Intestinal location- and age-specific variation of intraepithelial T lymphocytes and mucosal microbiota in pigs.

Intraepithelial T lymphocytes (T-IELs) are T cells located within the epithelium that provide a critical line of immune defense in the intestinal tract. In pigs, T-IEL abundances and phenotypes are used to infer putative T-IEL functions and vary by intestinal location and age, though investigations regarding porcine T-IELs are relatively limited. In this study, we expand on analyses of porcine intestinal T-IELs to include additional phenotypic designations not previously recognized in pigs. We describe non-conventional CD8α+CD8ß- αß T-IELs that were most prevalent in the distal intestinal tract and primarily CD16+CD27-, a phenotype suggestive of innate-like activation and an activated cell state. Additional T-IEL populations included CD8α+CD8ß+ αß, CD2+CD8α+ γδ, and CD2+CD8α- γδ T-IELs, with increasing proportions of CD16+CD27- phenotype in the distal intestine. Thus, putative non-conventional, activated T-IELs were most abundant in the distal intestine within multiple γδ and αß T-IEL populations. A comparison of T-IEL and respective mucosal microbial community structures across jejunum, ileum, and cecum of 5- and 7-week-old pigs revealed largest community differences were tissue-dependent for both T-IELs and the microbiota. Between 5 and 7 weeks of age, the largest shifts in microbial community compositions occurred in the large intestine, while the largest shifts in T-IEL communities were in the small intestine. Therefore, results indicate different rates of community maturation and stabilization for porcine T-IELs and the mucosal microbiota for proximal versus distal intestinal locations between 5 and 7 weeks of age. Collectively, data emphasize the intestinal tract as a site of location- and age-specific T-IEL and microbial communities that have important implications for understanding intestinal health in pigs.

Strategies to manage barn feed supply to prolong and hold late finishing pigs during a supply chain disruption.

The U.S. pork production system is sensitive to supply chain disruptions, including those that can create challenges of feed delivery and feed management during the event of a foreign animal disease outbreak. Therefore, the objective was to evaluate feeding strategies during a prolonged feed availability shortage in group-housed finishing pigs and assess the impacts on pig performance. A total of 1,407 mixed-sex pigs (92 ± 11 kg BW) were randomly allocated to one of five treatments across 60 pens (N = 12 pens per treatment, 22 pigs per pen) and were blocked by initial body weight (BW) within the replicate, over a 21-d test period. Treatments were fed for 14 d (P1), and thereafter all pens returned to ad libitum access to a standard commercial diet for 7 d (P2). Treatments included: 1) Pens fed ad libitum (CON); 2) Pens fed at 1.45X ME maintenance requirement daily of CON diet (1.45X); 3) Pens fed 2X ME maintenance requirement daily of CON diet (2X); 4) Tightened feeders to the lowest setting, fed ad libitum of CON diet (CF); and 5) whole corn kernels, fed ad libitum (WC). P1 and P2 BW and feed disappearance were recorded to calculate ADG, ADFI, and G:F. Data were analyzed with pen as the experimental unit and least-squares means values reported by treatment. Compared to CON, pens fed 1.45X, 2X, CF, and WC treatments had significantly reduced P1 ADG (1.09 vs. 0.02, 0.34, 0.72, 0.41 kg/d, respectively), ADFI (3.21 vs. 1.42, 1.90, 2.49, 2.40 kg/d, respectively) and G:F (P < 0.05). During P2, ADG and G:F were increased (P < 0.05) compared to CON across all treatments. However, ADFI increased only in the 2X, CF, and WC diet from the CON (P < 0.05). Overall (days 0 to 21), all strategies attenuated BW, ADG, and ADFI (P < 0.01) compared to CON. However, G:F was only reduced (P < 0.01) in 1.45X and WC, but not 2X and CF (P > 0.05) compared to CON. In conclusion, all strategies explored could extend feed budgets. Even though these strategies were successful, increased BW variability was reported with more restrictive strategies. Further, adverse pig behaviors and welfare implications needs to be considered in adopting any restrictive feeding strategy.

Functions of manganese in reproduction.

Manganese (Mn) is the twelfth most abundant element in the earth's crust and is widely distributed throughout the surface of the planet, naturally occurring in rocks, soil, water, and food. As an essential trace mineral in diets, Mn is required for a variety of metabolic functions including skeletal system development, energy metabolism, enzyme activation, nervous system function, immune system function, and reproductive hormone function. Manganese has effects on reproductive hormone function as a cofactor for enzymes necessary for cholesterol synthesis. Production of steroid hormones necessary for reproduction is dependent on the availability of cholesterol as a precursor. There is also evidence that Mn has effects on reproduction due to actions at the hypothalamus. Because Mn is used for manufacturing of steel, recent research has focused on the effects of Mn toxicity as a result of occupational endeavors rather than evaluating the optimal Mn inclusion rate for mammalian growth and development, reproductive function, immune function, etc. The objective of this review is to address the functions of Mn in reproduction of animals but there is also a focus on other areas of mammalian biology affected by Mn functions, with an emphasis on domestic swine (Sus scrofa).

Acute infection with Brachyspira hyodysenteriae affects mucin expression, glycosylation, and fecal MUC5AC.

Introduction: Infection with strongly ß-hemolytic strains of Brachyspira hyodysenteriae leads to swine dysentery (SD), a production-limiting disease that causes mucohemorrhagic diarrhea and typhlocolitis in pigs. This pathogen has strong chemotactic activity toward mucin, and infected pigs often have a disorganized mucus layer and marked de novo expression of MUC5AC, which is not constitutively expressed in the colon. It has been shown that fucose is chemoattractant for B. hyodysenteriae, and a highly fermentable fiber diet can mitigate and delay the onset of SD. Methods: We used lectins targeting sialic acids in α-2,6 or α-2,3 linkages, N-acetylglucosamine (GlcNAc), α-linked L-fucose, and an immunohistochemical stain targeting N-glycolylneuraminic acid (NeuGc) to investigate the local expression of these mucin glycans in colonic tissues of pigs with acute SD. We used a commercial enzyme-linked immunosorbent assay (ELISA) to quantify fecal MUC5AC in infected pigs and assess its potential as a diagnostic monitoring tool and RNA in situ hybridization to detect IL-17A in the colonic mucosa. Results: Colonic mucin glycosylation during SD has an overall increase in fucose, a spatially different distribution of GlcNAc with more expression within the crypt lumens of the upper colonic mucosa, and decreased expression or a decreased trend of sialic acids in α-2,6 or α-2,3 linkages, and NeuGc compared to the controls. The degree of increased fucosylation was less in the colonic mucosa of pigs with SD and fed the highly fermentable fiber diet. There was a significant increase in MUC5AC in fecal and colonic samples of pigs with SD at the endpoint compared to the controls, but the predictive value for disease progression was limited. Discussion: Fucosylation and the impact of dietary fiber may play important roles in the pathogenesis of SD. The lack of predictive value for fecal MUC5AC quantification by ELISA is possibly due to the presence of other non-colonic sources of MUC5AC in the feces. The moderate correlation between IL-17A, neutrophils and MUC5AC confirms its immunoregulatory and mucin stimulatory role. Our study characterizes local alteration of mucin glycosylation in the colonic mucosa of pigs with SD after B. hyodysenteriae infection and may provide insight into host-pathogen interaction.

The Use of Attractants to Stimulate Neonatal Piglet Interest in Rope Enrichment.

In the United States swine industry, preweaning mortality represents the highest mortality rate of any production phase, nearly half attributed to crushing. The overarching aim of this study was to determine if enrichment ropes would entice neonatal piglets away from the sow and reduce preweaning mortality. Rope enrichments were provided to 161 piglets from 26 sows after farrowing. Ropes were dipped in sunflower oil (n = 7), semiochemical (n = 8), or milky cheese (n = 11). Piglet purposeful rope investigations, weight gain, and mortality were recorded. On Day 2, 75% of piglets touched the enrichment at least once, and frequency ranged from 1 to 21 investigations across all treatments. Frequency (p = 0.20) and duration (p = 0.21) of investigations were not affected by treatment. Preweaning litter average weight gain did not differ between treatments (p = 0.71). MC (milky cheese) piglets had the lowest percent mortality when the enrichment ropes were present (Days 2 to 5, p = 0.01), and SC (semiochemical) piglets had the lowest percent mortality after the enrichment ropes were removed (Days 6 to weaning, p < 0.0001). This proof-of-concept study highlights the potential value of neonatal piglet environmental enrichment.

Evaluation of digestively resistant or soluble fibers, short- and medium-chain fatty acids, trace minerals, and antibiotics in nonchallenged nursery pigs on performance, digestibility, and intestinal integrity.

Three experiments (EXP) were conducted to determine the effect of feed additives on performance, intestinal integrity, gastrointestinal volatile fatty acids (VFA), and energy and nutrient digestion in nonchallenged nursery pigs. In EXP 1, 480 pigs (6.36-kg body weight, BW) were placed into 96 pens with 5 pigs/pen, and allotted to 1 of 10 dietary treatments: 1) negative control containing no feed additive (NC), 2) NC + 44 mg chlortetracycline and 38.5 mg tiamulin/kg diet (CTsb), 3) NC + 5% resistant potato starch (RSpo), 4) NC + 5% soluble corn fiber (SCF), 5) NC + 5% sugar beet pulp (SBP), 6) NC + 0.30% fatty acid mix (FAM), 7) NC + 0.10% phytogenic blend of essential oils and flavoring compounds (PHY), 8) NC + 50 mg Cu and 1,600 mg zinc oxide/kg diet (CuZn), 9) NC + 5% resistant corn starch (RScn), and 10) NC + 0.05% ß-glucan (BG) for 28 d. There was no impact of dietary treatment on BW gain or feed intake (P ≥ 0.22). Pigs fed diets containing SCF, CTsb, and RSpo resulted in microbial community differences compared to pigs fed the NC (P < 0.05). In EXP 2, 48 barrows (12.8 kg BW) were selected at the end of EXP 1 and fed the same dietary treatments they had previously received: 1) NC, 2) NC + 5% RScn, 3) NC + 5% SCF, and 4) NC + FAM for 8 d. There was no effect of feeding diets containing RScn, SCF, or FAM on in vivo intestinal permeability (P ≤ 0.21). Ileal or colon pH, concentrations of VFA did not differ due to dietary treatment (P ≥ 0.36), but pigs fed diets containing FAM resulted in a greater butyric acid concentration in the cecum compared to pigs fed the NC (P ≤ 0.05). In EXP 3, 156 pigs (6.11 kg BW) were placed into 52 pens with 3 pigs/pen and allotted to 1 of 4 dietary treatments arranged in a factorial manner: 1) NC, 2) NC + 5% RSpo, 3) NC + 0.30% FAM, and 4) NC + 5% RSpo + 0.30% FAM for 24 d. Feeding pigs diets containing RSpo did not affect BW gain (P = 0.91) while pigs fed diets containing FAM grew improved BW gain (P = 0.09). Colonic butyric acid concentrations were greater in pigs fed diets containing RSpo (P = 0.03), while pigs fed diets containing FAM exhibited reduced total VFA concentrations (P = 0.11). The results indicate that supplementing diets with digestively resistant but fermentable fibers, short- and medium-chain fatty acids, or antibiotics do not have a consistent effect, positive or negative, on markers of intestinal integrity or barrier function, intestinal VFA patterns, ATTD of energy and nutrients, or on pig performance.

In-feed antimicrobials have been an important technology in swine production for protecting health and supporting growth. However, with legislative restrictions on the use of most antibiotics for growth promotion, research is needed to evaluate in-feed additives in replacing this growth promoting technology. Thus, strategies to enhance energy and nutrient digestibility, intestinal function and integrity, gastrointestinal volatile fatty acid concentrations, and microbial ecology in nursery pigs are desirable targets. The results of the three experiments conducted herein do not indicate that supplementing diets with digestively resistant but fermentable fibers, short-medium-chain fatty acids, or antibiotics have a consistent positive or negative effect on markers of intestinal integrity or barrier function, VFA patterns (ileal, cecal, or colon), ATTD of energy and nutrients, or pig performance.

Highly Fermentable Fiber Alters Fecal Microbiota and Mitigates Swine Dysentery Induced by Brachyspira hyodysenteriae.

Brachyspira hyodysenteriae is an etiological agent of swine dysentery (SD). Diet fermentability plays a role in development of SD, but the mechanism(s) of action are largely unknown. Thus, this study aimed to determine whether replacing lowly fermentable fiber with highly fermentable fiber would mitigate a 42 d B. hyodysenteriae challenge. Thirty-nine barrows were allocated to dietary treatment groups: (1) 20% corn distillers dried grain with solubles (DDGS), 0% beet pulp (BP) or resistant starch (RS; lowly fermentable fiber (LFF)); (2) 10% DDGS, 5% BP, 5% RS (medium fermentable fiber (MFF)); and (3) 0% DDGS, 10% BP, 10% RS (highly fermentable fiber (HFF)). On day post inoculation 0, pigs were inoculated with B. hyodysenteriae. Overall, 85% LFF pigs developed clinical SD, 46% of MFF pigs developed SD, and 15% of HFF pigs developed SD (p < 0.05). Overall average daily gain (ADG) differed among all treatments (p < 0.001), with LFF pigs having the lowest ADG. For HFF pigs, ADG was 37% greater than LFF pigs (p < 0.001) and 19% greater than MFF pigs (p = 0.037). The LFF diet had greater relative abundance of Shuttleworthia and Ruminococcus torques. Further, microbiota of pigs that developed SD had enriched Prevotellaceae. Collectively, replacing DDGS with highly fermentable fiber reduced clinical SD, improved performance, and modulated fecal microbiota during B. hyodysenteriae challenge.

Evaluation of increased fiber, decreased amino acids, or decreased electrolyte balance as dietary approaches to slow finishing pig growth rates.

In swine production, pig movement restrictions or packing plant closures may create the need to slow growth rates of finishing pigs to ensure they remain at a marketable body weight when packing plant access is restored. Although dietary formulations can be successful at slowing pig growth, precision is needed regarding how to best formulate diets to achieve growth rate reductions. Thus, the objective was to evaluate three dietary experimental approaches aimed at slowing growth rates in finishing pigs. These approaches consisted of either increasing neutral detergent fiber (NDF), reducing essential amino acids, or reducing the dietary electrolyte balance through the addition of acidogenic salts. A total of 94 mixed-sex pigs (72.4 ± 11.2 kg BW) across two replicates were individually penned and assigned to 1 of 8 dietary treatments (n = 11-12 pigs/treatment): 1) Control diet representative of a typical corn-soybean meal-based finisher diet (CON); 2) diet containing 15% NDF from soybean hulls (15% NDF); 3) diet containing 20% NDF from soybean hulls (20% NDF); 4) diet containing 25% NDF from soybean hulls (25% NDF); 5) diet formulated as per CON but with 50% of the soybean meal replaced with corn (89% Corn); 6) diet containing 97% corn and no soybean meal or synthetic amino acids (97% Corn); 7) diet containing 2% anhydrous calcium chloride (2% CaCl2); and 8) diet containing 4% anhydrous calcium chloride (4% CaCl2). Over 28 d, pig body weights and performance were recorded weekly. At d 28, all pigs were ultrasound scanned and switched to the CON diet to evaluate compensatory gain from d 28 to 35. Overall, increased NDF did not impact any growth performance parameter (P > 0.05). Amino acid restriction reduced average daily gain (ADG), average daily feed intake (ADFI), and gain:feed (G:F) linearly (linear P < 0.001). Similarly, ADG, ADFI, and G:F were linearly reduced with increased CaCl2 inclusion (linear P < 0.001). ADG differed during the compensatory gain period (P < 0.001), with 4% CaCl2-fed pigs having a 47% increase in ADG compared with CON-fed pigs. Conversely, 15% and 25% NDF-fed pigs had reduced ADG compared with CON-fed pigs during the compensatory gain period. Gain efficiency differed from day 28 to 35 (P < 0.001), with 4% CaCl2-fed pigs having a 36% increase in G:F compared with CON-fed pigs. Altogether, these data demonstrate that both amino acid restriction and CaCl2 inclusion are effective at slowing pig growth, albeit at greater inclusion rates.

Epithelial-mesenchymal transition of absorptive enterocytes and depletion of Peyer's patch M cells after PEDV infection.

This study focused on intestinal restitution including phenotype switching of absorptive enterocytes and the abundance of different enterocyte subtypes in weaned pigs after porcine epidemic diarrhea virus (PEDV) infection. At 10 days post-PEDV-inoculation, the ratio of villus height to crypt depth in both jejunum and ileum had restored, and the PEDV antigen was not detectable. However, enterocytes at the villus tips revealed epithelial-mesenchymal transition (EMT) in the jejunum in which E-cadherin expression decreased while expression of N-cadherin, vimentin, and Snail increased. Additionally, there was reduced expression of actin in microvilli and Zonula occludens-1 (ZO-1) in tight junctions. Moreover, the protein concentration of transforming growth factor ß1 (TGFß1), which mediates EMT and cytoskeleton alteration, was increased. We also found a decreased number of Peyer's patch M cells in the ileum. These results reveal incomplete restitution of enterocytes in the jejunum and potentially impaired immune surveillance in the ileum after PEDV infection.

Nutritional approaches to slow late finishing pig growth: implications on carcass composition and pork quality.

Although pork producers typically aim to optimize growth rates, occasionally it is necessary to slow growth, such as when harvest facility capacity is limited. In finishing pigs, numerous dietary strategies can be used to slow growth so pigs are at optimal slaughter body weights when harvest facility capacity and/or access is restored. However, the impact of these diets on pork carcass quality is largely unknown. Thus, this study aimed to evaluate the efficacy of dietary strategies to slow growth in late finishing pigs and evaluate their effects on carcass composition and pork quality. Mixed-sex pigs (n = 897; 125 ± 2 kg BW) were randomly allotted across 48 pens and assigned to 1 of 6 dietary treatments (n = 8 pens/treatment): (1) Control diet representative of a typical finisher diet (CON); (2) diet containing 3% calcium chloride (CaCl2); (3) diet containing 97% corn and no soybean meal (Corn); (4) diet deficient in isoleucine (LowIle); (5) diet containing 15% neutral detergent fiber (NDF) from soybean hulls (15% NDF); and (6) diet containing 20% NDF from soybean hulls (20% NDF). Over 42 d, pen body weights and feed disappearance were collected. Pigs were harvested in 3 groups (14, 28, and 42 d on feed) and carcass data collected. From the harvest group, 1 loin was collected from 120 randomly selected carcasses (20 loins/treatment) to evaluate pork quality traits. Overall, ADG was reduced in CaCl2, Corn, and 20% NDF pigs compared with CON pigs (P < 0.001). However, ADFI was only reduced in CaCl2 and 20% NDF pigs compared with CON (P < 0.001). Feed efficiency was reduced in CaCl2 and Corn pigs compared with CON (P < 0.001). Hot carcass weights were reduced in CaCl2 pigs at all harvest dates (P < 0.001) and were reduced in Corn and 20% NDF pigs at days 28 and 42 compared with CON pigs (P < 0.001). In general, CaCl2 and 20% NDF diets resulted in leaner carcasses, whereas the Corn diet increased backfat by 42 d on test (P < 0.05). Loin pH was reduced and star probe increased in CaCl2 pigs compared with CON pigs (P < 0.05); no treatments differed from CON pigs regarding drip loss, cook loss, color, firmness, or marbling (P ≥ 0.117). Overall, these data indicate that several dietary strategies can slow finishing pig growth without evidence of behavioral vices. However, changes to carcass composition and quality were also observed, indicating quality should be taken into consideration when choosing diets to slow growth.