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.

Effects of ß-glucan on Salmonella enterica serovar Typhimurium swine colonization and microbiota alterations.

BACKGROUND: The 2017 Veterinary Feed Directive eliminated the use of medically important antibiotics for growth promotion of food animals; thus, alternative growth promoters are highly desirable by food animal producers to enhance animal health and reduce pathogen colonization, including the human foodborne pathogen Salmonella. ß(1-3)(1-6)-D-glucan (ß-glucan) is a soluble fiber with prebiotic characteristics; it has been shown to modulate immune and intestinal functions that strengthen swine resistance to health challenges such as bacterial infections when supplemented in the diets of growing pigs. The current study evaluated the effects of a ß-glucan product on gut microbial community structure as well as Salmonella shedding and intestinal colonization. RESULTS: Five-week-old pigs were fed a ß-glucan amended diet at 500 g/ton (n = 13) or a non-amended control diet (n = 14) for three weeks, followed by inoculation of the 27 pigs with 1 × 109 colony forming units of Salmonella enterica serovar Typhimurium strain UK1. While remaining on the respective diets, fecal samples collected at 2, 4, 7, and 16 days post-inoculation (dpi) were similar for Salmonella shedding counts between the two diets. At 16 dpi, Salmonella counts were significantly lower in the cecal contents of the ß-glucan-fed pigs (P = 0.0339) and a trend towards a reduction was observed in the Peyer's patches region of the ileum (P = 0.0790) compared to the control pigs. Pigs fed ß-glucan for three weeks exhibited an increase in members of the Clostridia class in their fecal microbial communities, and after inoculation with Salmonella, several potentially beneficial microorganisms were enriched in the microbiota of ß-glucan-fed pigs (Lactobacillus, Ruminococcaceae, Prevotellaceae, Veillonellaceae, Bifidobacterium and Olsenella). CONCLUSION: Administration of ß-glucan altered the swine gut microbiome and reduced Salmonella colonization in the cecal contents.

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.

Feeding thermally processed spray-dried egg whites, singly or in combination with 15-acetyldeoxynivalenol or peroxidized soybean oil on growth performance, digestibility, intestinal morphology, and oxidative status in nursery pigs.

Two experiments (EXP) determined the susceptibility of spray-dried egg white (SDEW) to oxidation (heating at 100 °C for 72 h; thermally processed, TP) and whether feeding TP-SDEW, 15-acetyldeoxynivalenol (15-ADON), or peroxidized soybean oil (PSO), singularly or in combination, would affect pig performance, intestinal morphology, digestibility, and markers of oxidative stress in nursery pigs. In EXP 1, 32 pigs (7.14 kg body weight, BW) were placed individually into pens and fed diets containing either 12% SDEW, 6% TP-SDEW plus 6% SDEW, or 12% TP-SDEW. Performance was measured at the end of the 24-d feeding period with biological samples harvested following euthanasia. In EXP 2, 64 pigs (10.6 kg BW) were placed individually into pens and fed diets containing 7.5% soybean oil or PSO, 10% SDEW or TP-SDEW, and diets without or with 3 mg 15-ADON/kg diet in a 2 × 2 × 2 factorial arrangement. Performance was measured at the end of the 28-d feeding period with biological samples harvested following euthanasia. In EXP 1, dietary treatment did not affect pig performance, apparent ileal digestibility of amino acids (AAs), apparent total tract digestibility (ATTD) of gross energy (GE) or nitrogen (N), ileal crypt depth, or villi height:crypt depth ratio (P > 0.05). The effects of feeding TP-SDEW on protein damage in the plasma and liver (P < 0.05) were variable. In EXP 2, there were no three-way interactions and only one two-way interactions among dietary treatments on parameters evaluated. There was no effect of feeding TP-SDEW on ATTD of GE or N, intestinal morphology, or on oxidative markers in the plasma, liver, or ileum (P > 0.05). There was no effect of feeding diets containing added 15-ADON on ATTD of GE, ileal AA digestibility, intestinal morphology, oxidative markers in the plasma, liver, or ileum, or pig performance (P > 0.05). Feeding pigs diets containing PSO resulted in reduced ATTD of GE and N, plasma vitamin E concentration, and pig performance (P < 0.01) but did not affect intestinal morphology or oxidative markers in the liver or ileum (P > 0.05). In conclusion, it was difficult to induce protein oxidation in SDEW and when achieved there were limited effects on performance, digestibility, intestinal morphology, and oxidative status. Furthermore, singly adding 15-A-DON to a diet had no effect on the animal. At last, adding PSO reduces animal performance, but has limited effect on digestibility, intestinal morphology, and oxidative status in nursery pigs.

Swine can be exposed to a variety of nutritional stressors that can affect their well-being and productivity. Three stressors of concern include grains with naturally occurring mycotoxins, oxidized proteins in feedstuffs due to overheating during processing, or lipids that have been damaged by excessive heating. Experiments were conducted to determine how susceptible a previously processed feedstuff was to protein oxidation and whether feeding mycotoxins, oxidized protein, or peroxidized soybean oil would affect growth performance, intestinal morphology, digestibility, and markers of oxidative stress in nursery pigs. Results indicate it was difficult to induce protein oxidation in previously processed protein by heating in a forced air oven, and if some protein oxidation did occur, there is limited effects on growth performance, digestibility, intestinal morphology, and oxidative status in nursery pigs. The data also indicated that adding an isolated mycotoxin was difficult to ensure proper mixing from which to analyze the complete diet from which to conduct animal research. At last, the data show that adding soybean oil that has been thermally processed to contain high concentrations of aldehydes will result in a dramatic reduction in animal performance, but has limited effects on digestibility, intestinal morphology, and oxidative status in nursery pigs.

Effect of dietary ß-glucan on intestinal microbial diversity and Salmonella vaccine immunogenicity and efficacy in pigs.

Alternatives to antibiotics to improve animal performance, limit the negative impact of infectious disease, and/or reduce colonization with foodborne pathogens is a major focus of animal agricultural research. ß-glucans, a generally-recognized-as-safe (GRAS) product derived from various sources, are used in swine and can serve as both a prebiotic and/or stimulant of the immune system given the expression of ß-glucan receptors on immune cells. When supplied in the diet of nursery pigs, it is unclear how dietary additives, particularly those known to modulate immune status, impact immunogenicity and efficacy of mucosal-delivered vaccines. Salmonellosis is one of the most common bacterial foodborne infections in the United States, and consumption of contaminated pork is a major source of human infection. Reduction of foodborne Salmonella in pigs via vaccination is one strategy to reduce contamination risk and subsequently reduce human disease. We examined the ability of dietary ß-glucan to modulate fecal microbial diversity, and immunogenicity and efficacy of a mucosally-delivered, live-attenuated Salmonella vaccine during the nursery period. While dietaryß-glucan did modulate fecal alpha diversity, it did not alter the induction of peripheral Salmonella-specific IFN-γ secreting Tcells or Salmonella-specific IgA in oral fluids. In addition, vaccination reduced Salmonella enterica serovar Typhimurium fecal shedding and tissue colonization. Overall, addition of ß-glucan to the nursery diet of pigs impacted the microbiota but did not alter mucosal vaccine immunogenicity and efficacy.

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.

Effects of feeding variable levels of mycotoxins with or without a mitigation strategy on growth performance, gut permeability, and oxidative biomarkers in nursery pigs.

The objectives were to determine how high levels (> 2.5 mg/kg diet) of deoxynivalenol (DON), in conjunction with other naturally occurring mycotoxins (MTX) would impact growth, intestinal integrity, and oxidative status, with or without a mitigation strategy, in nursery pigs. One-hundred and five pigs (5.5 ± 0.52 kg) were randomly allotted to 35 pens and fed dietary treatments for 45 d. Treatments were factorially arranged with the inclusion of MTX being low (L-MTX; < 1 mg/kg diet) or high (H-MTX; > 2.5 mg/kg diet) in combination with no mitigation strategy or the inclusion of a mitigation strategy (Biofix® Plus, BPL; 1.5 mg/kg diet). There was no interaction between MTX level and BPL inclusion on average daily gain (ADG) or gain to feed ratio (GF), (P > 0.10). Compared to pigs fed diets containing L-MTX, feeding pigs diets containing H-MTX decreased ADG and GF (P < 0.05). The addition of BPL had no effect on ADG (P > 0.10), but improved GF (P = 0.09). There was an interaction between MTX and BPL on average daily feed intake (ADFI), where the addition of BPL had no effect on ADFI of pigs fed L-MTX diets but improved ADFI of pigs fed H-MTX diets (P = 0.09). An interaction was detected between MTX and BPL on protein oxidation as measured by plasma protein carbonyls (PC, P = 0.01), where the inclusion of BPL decreased plasma PC in pigs fed H-MTX diets to a greater extent than pigs fed the L-MTX diets. There was no interaction between MTX and BPL, or an effect of MTX or BPL on DNA damage as measured by 8-hydroxy-2'dexoxyguanosine (P > 0.10). There was no interaction between MTX and BPL, or a BPL effect on lipid damage as measured by thiobarbituic acid reactive substances (TBARS, P > 0.10), but pigs fed diets containing H-MTX exhibited lower concentrations of plasma TBARS (P = 0.07) compared to pigs fed L-MTX diets. There was no interaction between MTX and BPL, or an effect of MTX or BPL on plasma lactulose and mannitol ratio as a measure of intestinal permeability (P > 0.10). In conclusion, feeding H-MTX decreased ADG and GF, decreased plasma TBARS, but did not affect plasma 8-hydroxy-2'dexoxyguanosine or plasma LM ratio. The inclusion of a mitigation strategy improved ADFI when pigs were fed H-MTX diets and improved GF regardless of MTX level. Addition of a mitigation strategy also reduced plasma protein damage but did not affect indicators of DNA or lipid damage or affect gastrointestinal integrity.

Short Chain Fatty Acids and Bacterial Taxa Associated with Reduced Salmonella enterica serovar I 4,[5],12:i:- Shedding in Swine Fed a Diet Supplemented with Resistant Potato Starch.

Salmonella enterica serovar I 4,[5],12:i:- is a foodborne pathogen of concern because many isolates are multidrug-resistant (resistant to ≥3 antimicrobial classes) and metal tolerant. In this study, three in-feed additives were individually tested for their ability to reduce Salmonella I 4,[5],12:i:- shedding in swine: resistant potato starch (RPS), high amylose corn starch, and a fatty acid blend, compared with a standard control diet over 21 days. Only RPS-fed pigs exhibited a reduction in Salmonella fecal shedding, different bacterial community compositions, and different cecal short chain fatty acid (SCFA) profiles relative to control animals. Within the RPS treatment group, pigs shedding the least Salmonella tended to have greater cecal concentrations of butyrate, valerate, caproate, and succinate. Additionally, among RPS-fed pigs, several bacterial taxa (Prevotella_7, Olsenella, and Bifidobacterium, and others) exhibited negative relationships between their abundances of and the amount of Salmonella in the feces of their hosts. Many of these same taxa also had significant positive associations with cecal concentrations of butyrate, valerate, caproate, even though they are not known to produce these SCFAs. Together, these data suggest the RPS-associated reduction in Salmonella shedding may be dependent on the establishment of bacterial cross feeding interactions that result in the production of certain SCFAs. However, directly feeding a fatty acid mix did not replicate the effect. RPS supplementation could be an effective means to reduce multidrug-resistant (MDR) S. enterica serovar I 4,[5],12:i:- in swine, provided appropriate bacterial communities are present in the gut. IMPORTANCE Prebiotics, such as resistant potato starch (RPS), are types of food that help to support beneficial bacteria and their activities in the intestines. Salmonella enterica serovar I 4,[5],12:i:- is a foodborne pathogen that commonly resides in the intestines of pigs without disease, but can make humans sick if unintentionally consumed. Here we show that in Salmonella inoculated pigs, feeding them a diet containing RPS altered the colonization and activity of certain beneficial bacteria in a way that reduced the amount of Salmonella in their feces. Additionally, within those fed RPS, swine with higher abundance of these types of beneficial bacteria had less Salmonella I 4,[5],12:i:- in their feces. This work illustrates likely synergy between the prebiotic RPS and the presence of certain gut microorganisms to reduce the amount of Salmonella in the feces of pigs and therefore reduce the risk that humans will become ill with MDR Salmonella serovar I 4,[5],12:i:-.

Feeding oxidized chicken byproduct meal impacts digestibility more than performance and oxidative status in nursery pigs.

Rendered products from the meat industry provide quality proteins in diets for companion animals. These proteins are exposed to extreme temperatures during processing leading to the potential for decreased diet digestibility and subsequent growth performance. While this would impact production efficiency in livestock species, oxidized ingredients in companion animal diets may impact health and longevity. The objective of this study was to determine the extent to which a feedstuff containing oxidized protein and lipid affect diet digestibility, growth performance, and oxidative stress in nursery pigs. A total of 56 male pigs (21 d of age, initial body weight 5.51 ± 0.65 kg) were randomly assigned to one of the four dietary treatments in a 2 × 2 factorial arrangement with two levels of heat and two levels of antioxidant (AOX). Diets were fed for 35 d and growth performance was measured, while total tract digestibility and nitrogen (N) balance was determined during the trial on day 18-20. Blood plasma was collected on day 34 and jejunum, colon, and liver tissues were collected on day 35 to analyze for markers of oxidative stress. Average daily feed intake (ADFI) was reduced in pigs fed diets without AOXs (P = 0.02). Additionally, pigs consuming diets containing heated chicken byproduct (CBP) meal had decreased gain:feed (GF; P = 0.02). There was an interaction between heat and AOX (P = 0.02) where heating CBP reduced N digestibility in the presence of an AOX but did not have an impact when AOX was not present. The removal of AOX resulted in reduced GE digestibility (P < 0.01). Dry matter (P < 0.01), ash (P < 0.01), and protein (P < 0.01) digestibility were reduced (P < 0.01) as a result of heating. Furthermore, heating (P =0.01) as well as absence of AOX (P =0.01) resulted in reduced digestible energy. No difference was detected in N retention suggesting that oxidation reduces digestibility but has no impact on N utilization. This is supported by the fact that systemic oxidative stress was not consistently affected by heating or AOX inclusion. These results suggest that feeding pigs CBP containing oxidized proteins and lipids did not induce oxidative stress. However, feeding young pigs CBP containing oxidized proteins and lipids did result in reduced energy and nutrient digestibility as well as negatively affected feed efficiency. Because CBP is commonly used in companion animal diets, it is reasonable to revisit their impacts on those species.

Lipid Source and Peroxidation Status Alter Immune Cell Recruitment in Broiler Chicken Ileum.

BACKGROUND: Restaurant oil in poultry diets increases energy content, reduces production costs, and promotes sustainability within the food supply chain. However, variable oil composition and heating temperatures among restaurant oil sources can impact broiler chicken health due to heat-induced lipid modifications. OBJECTIVES: A 21-d experiment was conducted to evaluate ileal morphology, liver cytokine gene expression, and ileal immune cell populations in broilers fed control or peroxidized lipids with varying chain and saturation characteristics. METHODS: Day-old broilers were housed in battery cages (5 birds per cage) and fed diets containing 5% control or peroxidized oils. Eight diets were randomly assigned in a 4 × 2 factorial arrangement consisting of oil source (palm, soybean, flaxseed, or fish) and peroxidation status (control or peroxidized). At day 21, samples were collected for ileal histomorphology [villus height (VH), crypt depth (CrD), and the VH:CrD ratio], and liver cytokine expression (qPCR). Ileum cytokine expression and T-cell markers were analyzed by RNAscope in situ hybridization (ISH). Data were analyzed as a mixed model (SAS 9.4) with fixed effects of lipid source, peroxidation, and lipid × peroxidation interaction. RESULTS: CD3+ T-cells in the ileum decreased 16.2% due to peroxidation (P = 0.001) with 30.3% reductions observed in birds fed peroxidized flaxseed oil (P = 0.01). Peroxidation increased IL6+ and IL1B+ cells by 62.0% and 40.3%, respectively (P = 0.01). Soybean oil increased IFNG+ cells by 55.1% compared with palm oil, regardless of peroxidation status (P = 0.007). Lipid source and peroxidation did not alter ileal histomorphology or liver cytokine expression. CONCLUSIONS: Lipid peroxidation increased ileal IL1B and IL6 in broiler chickens, whereas soybean oil diets increased IFNG. Generally, peroxidation decreased overall CD3+ T-cell populations, suggesting impaired T-cell presence or recruitment. These results identify potential immunomodulatory lipid profiles in restaurant oil while supporting RNAscope-ISH as a method to describe avian tissue-level immune responses.

Influence of feeding thermally peroxidized lipids on growth performance, lipid digestibility, and oxidative status in nursery pigs.

Three experiments were conducted to evaluate oil source and peroxidation status (experiment 1) or peroxidized soybean oil (SO; experiments 2 and 3) on growth performance, oxidative stress, and digestibility of dietary ether extract (EE). In experiment 1, palm oil (PO), poultry fat (PF), canola oil (CO), and SO were evaluated, while in experiments 2 and 3, only SO was evaluated. Lipids were either an unheated control (CNT) or thermally processed at 90 °C for 72 hr, being added at 10%, 7.5%, or 3% of the diet in experiments 1, 2, and 3, respectively. In experiment 1, 288 pigs (body weight, BW, 6.1 kg) were fed 1 of 8 factorially arranged treatments with the first factor being lipid source (PO, PF, CO, and SO) and the second factor being peroxidation status (CNT or peroxidized). In experiment 2, 216 pigs (BW 5.8 kg) were fed 1 of 6 treatments consisting of 100%, 90%, 80%, 60%, 20%, and 0% CNT SO blended with 0%, 10%, 20%, 40%, 80%, and 100% peroxidized SO, respectively. In experiment 3, 72 pigs (BW 5.8 kg) were fed either CNT or peroxidized SO. Pigs were fed 21 d with feces collected on day 12 or 14 and pigs bled on day 12 blood collection. In experiment 1, an interaction between oil source and peroxidation status was observed for averaged daily gain (ADG) and average daily feed intake (ADFI; P = 0.10) which was due to no impact of feeding pigs peroxidized PO, PF, or SO on ADG or ADFI compared with feeding pigs CNT PO, PF, or SO, respectively; while pigs fed peroxidized CO resulted in reduced ADG and ADFI compared with pigs fed CNT CO. There was no interaction between oil source and peroxidation status, and no lipid source effect on gain to feed ratio (GF; P ≥ 0.84), but pigs fed the peroxidized lipids had a lower GF compared with pigs fed the CNT lipids (P = 0.09). In experiment 2, feeding pigs diets containing increasing levels of peroxidized SO resulted in reduced ADG (quadratic, P = 0.03), ADFI (linear, P = 0.01), and GF (quadratic, P = 0.01). In experiment 3, feeding peroxidized SO at 3% of the diet reduced ADG (P = 0.11) and ADFI (P = 0.13), with no observed change in GF (P = 0.62). Differences in plasma protein carbonyls, glutathione peroxidase, and vitamin E due to feeding peroxidized lipids were inconsistent across the 3 experiments. Digestibility of dietary EE was reduced in pigs fed peroxidized PO or SO (P = 0.01, experiment 1) and peroxidized SO in experiments 2 and 3 (P ≤ 0.02). In conclusion, the peroxidation status of dietary lipids consistently affects growth performance and EE digestibility but has a variable effect on measures of oxidative stress.

The effects of enzymatically treated soybean meal on growth performance and intestinal structure, barrier integrity, inflammation, oxidative status, and volatile fatty acid production of nursery pigs.

The objective of this experiment was to determine the impact of diets containing increasing amounts of enzymatically treated soybean meal (ESBM) but decreasing amounts of soybean meal (SBM) on growth performance, intestinal structure, and barrier integrity, inflammation, and oxidative status in weaned pigs. A total of 480 pigs [6.3 ± 1.2 kg body weight (BW)] were blocked by initial BW and pens (n = 12 per treatment) were randomly allotted to one of four dietary treatments. Diets were fed in three phases (days 0-14, 14-28, and 28-35) over a 35-d period. The four dietary treatments consisted of a negative control diet (NC): the NC with 7.0% ESBM (ESBM1), the NC with 14.0% ESBM (ESBM2), and the NC with 21.0% ESBM (ESBM3). Soybean meal was reduced proportionately in each treatment. In phase 2, ESBM inclusion was decreased by 50% (3.5%, 7.0%, and 10.5% ESBM, respectively); phase 3 was a common diet and contained no ESBM. Fecal score was visually ranked weekly using a four-point scale. Intestinal tissue, digesta, and blood samples were collected from 48 pigs (1 per pen) on day 10. Data were analyzed using PROC MIXED of SAS (9.4) with pen as the experimental unit; diet and block were considered fixed effects. Linear and quadratic contrasts were used to determine the effect of increasing ESBM. Overall, ESBM2 and ESBM3 decreased final BW, average daily gain, and average daily feed intake compared to NC and ESBM1 (diet, P < 0.05; linear, P < 0.05). Overall fecal score (diet, P < 0.05) and fecal dry matter (P < 0.05) were improved by feeding ESBM diets compared to NC. Volatile fatty acid (VFA) concentration of acetate, propionate, butyrate, and total VFA in ileal contents increased as ESBM inclusion increased (P < 0.05). Colonic VFA concentration was not impacted (P > 0.10). Total antioxidant capacity was increased by ESBM (P < 0.05). The concentration of mucosal interleukin-4 increased as the inclusion of ESBM increased (linear, P < 0.05). Messenger ribonucleic acid abundance of occludin and zonula-occludens-1 in ileal tissue was increased by ESBM1 or ESBM2 (P < 0.05). In conclusion, increasing the dietary levels of ESBM over 7% had a negative impact on nursery pig performance, but ESBM positively impacted fecal score. Feeding ESBM improved oxidative status and intestinal barrier integrity while increasing ileal VFA production but had minimal impact on intestinal inflammation or morphology. Further research is needed to determine the optimal inclusion level of ESBM.

The Role of Salmonella Genomic Island 4 in Metal Tolerance of Salmonella enterica Serovar I 4,[5],12:i:- Pork Outbreak Isolate USDA15WA-1.

Multidrug-resistant (MDR; resistance to >3 antimicrobial classes) Salmonella enterica serovar I 4,[5],12:i:- strains were linked to a 2015 foodborne outbreak from pork. Strain USDA15WA-1, associated with the outbreak, harbors an MDR module and the metal tolerance element Salmonella Genomic Island 4 (SGI-4). Characterization of SGI-4 revealed that conjugational transfer of SGI-4 resulted in the mobile genetic element (MGE) replicating as a plasmid or integrating into the chromosome. Tolerance to copper, arsenic, and antimony compounds was increased in Salmonella strains containing SGI-4 compared to strains lacking the MGE. Following Salmonella exposure to copper, RNA-seq transcriptional analysis demonstrated significant differential expression of diverse genes and pathways, including induction of at least 38 metal tolerance genes (copper, arsenic, silver, and mercury). Evaluation of swine administered elevated concentrations of zinc oxide (2000 mg/kg) and copper sulfate (200 mg/kg) as an antimicrobial feed additive (Zn+Cu) in their diet for four weeks prior to and three weeks post-inoculation with serovar I 4,[5],12:i:- indicated that Salmonella shedding levels declined at a slower rate in pigs receiving in-feed Zn+Cu compared to control pigs (no Zn+Cu). The presence of metal tolerance genes in MDR Salmonella serovar I 4,[5],12:i:- may provide benefits for environmental survival or swine colonization in metal-containing settings.

Dietary composition and particle size effects on swine manure characteristics and gas emissions.

Nutrients excreted from animals affect the nutritive value of manure as a soil amendment as well as the composition of gases emitted from manure storage facilities. There is a dearth of information, however, on how diet type in combination with dietary particle size affects nutrients deposited into manure storage facilities and how this affects manure composition and gas emissions. To fill this knowledge gap, an animal feeding trial was performed to evaluate potential interactive effects between feed particle size and diet composition on manure characteristics and manure-derived gaseous emissions. Forty-eight finishing pigs housed in individual metabolism crates that allowed for daily collection of urine and feces were fed diets differing in fiber content and particle size. Urine and feces were collected and stored in 446-L stainless steel containers for 49 d. There were no interactive effects between diet composition and feed particle size on any manure or gas emission parameter measured. In general, diets higher in fiber content increased manure nitrogen (N), carbon (C), and total volatile fatty acid (VFA) concentrations and increased manure VFA emissions but decreased manure ammonia emissions. Decreasing the particle size of the diet lowered manure N, C, VFAs, phenolics, and indole concentrations and decreased manure emissions of total VFAs. Neither diet composition nor particle size affected manure greenhouse gas emissions.

Identification of C9-C11 unsaturated aldehydes as prediction markers of growth and feed intake for non-ruminant animals fed oxidized soybean oil.

BACKGROUND: The benefits of using the oxidized oils from rendering and recycling as an economic source of lipids and energy in animal feed always coexist with the concerns that diverse degradation products in these oxidized oils can negatively affect animal health and performance. Therefore, the quality markers that predict growth performance could be useful when feeding oxidized oils to non-ruminants. However, the correlations between growth performance and chemical profiles of oxidized oils have not been well examined. In this study, six thermally oxidized soybean oils (OSOs) with a wide range of quality measures were prepared under different processing temperatures and processing durations, including 45 °C-336 h; 67.5 °C-168 h; 90 °C-84 h; 135 °C-42 h; 180 °C-21 h; and 225 °C-10.5 h. Broilers and nursery pigs were randomly assigned to diets containing either unheated control soybean oil or one of six OSOs. Animal performance was determined by measuring body weight gain, feed intake, and gain to feed ratio. The chemical profiles of OSOs were first evaluated by common indicative tests, including peroxide value, thiobarbituric acid reactive substances, p-anisidine value, free fatty acids, oxidized fatty acids, unsaponifiable matter, insoluble impurities, and moisture, and then analyzed by the liquid chromatography-mass spectrometry-based chemometric analysis. RESULTS: Among common quality indicators, p-anisidine value (AnV), which reflects the level of carbonyl compounds, had the greatest inverse correlation with the growth performance of both broilers and pigs, followed by free fatty acids and oxidized fatty acids. Among the 17 aldehydes identified in OSOs, C9-C11 alkenals, especially 2-decenal and 2-undecenal, had stronger inverse correlations (r < - 0.8) with animal performance compared to C5-C8 saturated alkanals, suggesting that chain length and unsaturation level affect the toxicity of aldehydes. CONCLUSIONS: As the major lipid oxidation products contributing to the AnV, individual C9-C11 unsaturated aldehydes in heavily-oxidized oils could function as effective prediction markers of growth and feed intake in feeding non-ruminants.

Impact of dietary oxidized protein on oxidative status and performance in growing pigs.

Rendered products from the meat industry can provide economical quality sources of proteins to the animal and feed industry. Similar to lipids, rendered proteins are susceptible to oxidation, yet the stability of these proteins is unclear. In addition, interest in understanding how oxidative stress can impact efficiency in production animals is increasing. Recent studies show that consumption of oxidized lipids can lead to a change in the oxidative status of the animal as well as decreases in production efficiency. To date, little is known about how consumption of oxidized proteins impacts oxidative status and growth performance. The objectives of this study were to determine if feeding diets high in oxidized protein to growing pigs would: 1) impact growth performance and 2) induce oxidative stress. Thirty pigs (42 d old; initial body weight [BW] 12.49 ± 1.45 kg) were randomly assigned to one of three dietary treatments with increasing levels of oxidized protein. Spray-dried bovine plasma was used as the protein source and was either unheated upon arrival, heated at 45 °C for 4 d, or heated at 100 °C for 3 d. Diets were fed for 19 d and growth performance was measured. Blood plasma (days 0 and 18), jejunum, colon, and liver tissues (day 19) were collected to analyze for markers of oxidative stress (e.g., protein oxidation, lipid oxidation, DNA damage, and glutathione peroxidase activity). Average daily gain (ADG;P < 0.01) and average daily feed intake (ADFI;P < 0.01) had a positive linear relationship to increased protein oxidation, but there was no effect on gain to feed ratio. Furthermore, protein (P = 0.03) and fat (P < 0.01) digestibility were reduced with increased protein oxidation in the diet. Crypt depth showed a positive linear relationship with dietary protein oxidation levels (P = 0.02). A trend was observed in liver samples where pigs fed the plasma heated to 45 °C had increased lipid oxidation compared with pigs fed the plasma either unheated or heated to 100 °C (P = 0.09). DNA damage in the jejunum tended to have a linear relationship with the dietary protein oxidation level (P = 0.07). Even though results suggest dietary oxidized protein did not induce oxidative stress during short-term feeding, differences in performance, gut morphology, and digestibility are likely a result of reduced protein availability.

Changes in markers of lipid oxidation and thermal treatment in feed-grade fats and oils.

BACKGROUND: Oxidized feed lipids have been shown to have detrimental effects on food animal growth and metabolism. The present study aimed to measure classes of lipid oxidation products (LOP) in feed-grade oils at temperatures representing production and storage conditions. RESULTS: There were significant oil type × time interactions in the accumulation of primary and secondary LOP. At 22.5 °C, peroxide value (PV), a marker for the primary phase of lipid oxidation, increased most in fish oil (FO), followed by tallow (TL), soybean oil (SO), linseed oil (LO) and modified algae oil (MAO), whereas palm oil (PO) showed no appreciable increase in PV. Secondary LOP, such as p-anisidine value, hexanal, 2,4,-decadienal, polymerized triacylglycerols and total polar compounds, increased only in FO. At 45 °C, FO and SO produced both primary and secondary LOP, whereas MAO, PO and TL had slower rates of PV increase and no secondary LOP. At 90 °C and 180 °C, all oils except for FO accumulated both primary and secondary LOP. CONCLUSIONS: Higher polyunsaturated fatty acid:saturated fatty acid oils and higher temperatures produced greater quantities of primary and secondary LOP. However, unrefined TL was more prone to oxidation at 22.5 °C than predicted, whereas LO was more stable than predicted, indicating that pro-oxidant and antioxidant compounds can markedly influence the rate of oxidation. Measuring both primary and secondary LOP will provide better information about the oxidative status of feed oils and provide better information about which classes of LOP are responsible for detrimental health effects in animals. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Lack of interactive effects between diet composition and acid addition with drying method on amino acid digestibility values in porcine ileal digesta.

Two experiments were conducted to determine the effect of oven drying (OD) or freeze drying (FD) on apparent ileal digestibility (AID) of AA in diets fed to pigs. In experiment 1, 15 barrows (88.4 ± 6.4 kg) were allotted to either a corn starch-soybean meal (CST), potato starch-soybean meal (PST), or corn-soybean meal (CSBM) diet, over 2 collection periods. Following collection, samples were pooled within pig and subdivided into either OD or FD, resulting in 10 observations per diet by drying-method combination. In experiment 2, 11 barrows (63.3 ± 3.8 kg) were fed a CST diet, and, following collection, samples were pooled within pig and subdivided and either adjusted to pH 4 or remain unadjusted. Subsets of these samples were then subdivided to be either FD or OD, resulting in 11 observations per pH level by drying-method combination. OD was accomplished by drying samples in a forced air oven at either 100 °C (experiment 1) or 75 °C (experiment 2). In experiment 1, there was no diet type by drying-method interactions noted for any of the AA (P > 0.10).OD resulted in a higher AID of AA compared with samples which were FD (P ≤ 0.10), for all AA except for Gly and Tyr. Averaged across all AA, AID of AA was 3.3% greater if the sample was OD compared with FD. Differences in AID of AA among the 3 diets were noted for all AA (P ≤ 0.07), except for Cys (P = 0.33), due to the fact that CST and PST diets only contained soybean meal (SBM) as an AA-providing ingredient while the CSBM diet contained both corn and SBM. Pigs fed the PST diet had greater SID for all AA compared with pigs fed the CST diet (P ≤ 0.05), except for His, Lys, Cys, and Glu. In experiment 2, there were no pH-adjustment by drying-method interactions noted on AID for any of the AA (P > 0.10). Adjusting ileal digesta to a pH of 4.0 had little effect on AID for most of the AA, except for a lowered AID of Arg, His, Lys, Trp, and Ser (P ≤ 0.10). OD resulted in a higher AID for all AA (P ≤ 0.09) except for Ile, Thr, Val, Ala, Asp, Glu, and Gly. Averaged across all AA, the increase in AID of AA was 1.7% greater if the sample was OD compared with FD. On average, OD of ileal digesta resulted in a 2.5% greater estimate of AID of AA compared with samples that were FD, and was not diet-, pH-, or AA-dependent. Because the majority of the data on AA digestibility are based on FD, a bias factor may be necessary to adjust AA digestibility data obtained on an OD-basis relative to an FD-basis for use in feed formulation.

Influence of feeding thermally peroxidized soybean oil on growth performance, digestibility, gut integrity, and oxidative stress in nursery pigs.

The objectives of the current experiments were to evaluate the effect of feeding soybean oil (SO) with different levels of peroxidation on lipid, N, and GE digestibility, gut integrity, oxidative stress, and growth performance in nursery pigs. Treatments consisted diets containing 10% fresh SO (22.5 °C) or thermally processed SO (45 °C for 288 h, 90 °C for 72 h, or 180 °C for 6 h), each with an air infusion of 15 L/min, with postprocessing peroxide values of 7.6, 11.5, 19.1, and 13.4 mEq/kg and p-anisidine values of 1.92, 6.29, 149, and 159, for the 22.5 °C, 45 °C, 90 °C and 180 °C processed SO, respectively. In experiment 1, 64 barrows (7.1 ± 0.9 kg initial BW) were randomly allotted into 2 rooms of 32 pens and individually fed their experimental diets for 21 d, with a fresh fecal sample collected on day 20 for determination of GE and lipid digestibility. In experiment 2, 56 barrows (BW 9.16 ± 1.56 kg) were placed into individual metabolism crates for assessment of GE, lipid, and N digestibility and N retention. Urinary lactulose to mannitol ratio was assessed to evaluate in vivo small intestinal integrity, and urine and plasma were collected to analyze for markers of oxidative stress. Pigs were subsequently euthanized to obtain liver weights and analyze the liver for markers of oxidative stress. In experiment 1, pigs fed the SO thermally processed at 90 °C had reduced ADG (P = 0.01) and ADFI (P = 0.04) compared to pigs fed the other SO treatment groups, with no differences noted among pigs fed the 22.5 °C, 45 °C, and 180 °C SO treatments. No effects of feeding thermally processing SO on dietary GE or lipid digestibility (P > 0.10) were noted in either experiment. In experiment 2, there was no dietary effect of feeding peroxidized SO on the DE:ME ratio, N digestibility, or N retained as a percent of N digested, on the urinary ratio of lactulose to mannitol, on serum, urinary, or liver thiobarbituric acid reactive substances, on plasma protein carbonyls, or on urinary or liver 8-OH-2dG (P > 0.10). In experiment 2, pigs fed the SO thermally processed at 90 °C had the greatest isoprostane concentrations in the serum (P ≤ 0.01) and urine (P ≤ 0.05) compared to pigs fed the unprocessed SO. These results indicate that the change in fatty acid composition and/or the presence of lipid peroxidation products in peroxidized SO may reduce ADG and ADFI in nursery pigs, but appears to have no impact on GE, lipid, or N digestibility, or gut permeability. These data suggest that the presence of lipid peroxidation products may affect certain markers of oxidative stress.

Amino acid composition and digestible amino acid content in animal protein by-product meals fed to growing pigs1.

An industry survey and animal experiment were conducted to evaluate the amino acid (AA) compositional variability and standardized ileal digestibility (SID) of AA in animal protein by-products fed to growing pigs. Animal protein by-product meals (212) were categorized into 8 groupings (blood meal, chicken by-product meal, chicken meal, feather meal, meat and bone meal, meat meal, poultry by-product meal, and poultry meal) and analyzed for total AA. Amino acid analysis among (e.g., Lys in blood meal averaged 9.20% compared with 2.31% for feather meal, DM basis) and within (e.g., Lys range of 1.54% in blood meal and 1.44% in feather meal, DM basis) the by-product classifications varied as expected, but on average the total AA values were similar to that reported in the literature. For the determination of the SID of AA, 15 barrows (average initial and final BW of 31.6 and 78.7 kg, respectively) were fitted with a T-cannula in the distal ileum and allotted to 15 diets over nine 7-d periods, resulting in 9 replications per diet. Pigs were fed a basal diet based on soybean meal and dehulled-degermed corn, 13 diets containing 17.5% animal protein by-product meal to partially replace a portion of the soybean meal and dehulled-degermed corn in the basal diet, or a N-free diet. Pigs were re-allotted to diets based on minimizing the number to times that the N-free diet would precede or follow feeding either of the blood or feather meal diets because of concern with inadequate diet consumption, as well as to prevent diets from being re-fed to the same pig during the next or subsequent periods. Values for the apparent ileal AA digestibility of each diet were determined, adjusted to SID based upon the endogenous AA losses determined by feeding the N-free diet, and the SID of AA in each animal protein by-product meal calculated using the difference procedure. The SID of AA varied among (e.g., SID of Lys averaged 91% in chicken meal but 47% in feather meal) and within (e.g., SID of Lys in three meat and bone meals was 80%, 71%, and 54%) the animal protein by-product meals, as would be expected and are consistent with variation reported in the literature. Overall, the data provide total AA composition for 212 samples and SID of AA for 13 samples of animal protein by-product meals, including data on their variability, which is critical for their use in feed formulation programs.