Effects of dietary protein content and crystalline amino acid supplementation patterns in low protein diets on intestinal bacteria and their metabolites in weaned pigs raised under Different sanitary conditions.

The objective of this experiment was to investigate the effects of dietary crude protein (CP) content and crystalline amino acids (CAA) supplementation patterns in low CP (LCP) diets on intestinal bacteria and their metabolites in weaned pigs raised under clean (CSC) or unclean sanitary conditions (USC). One hundred forty-four piglets (6.35 ±â€…0.63 kg) were assigned to one of six treatments in a 3 × 2 factorial arrangement based on CP content and sanitary conditions in a randomized complete block design to give eight replicates with three pigs per pen over a 21-d period. Diets consisted of a high CP (HCP; 21%) and two LCP (18%) diets supplemented with 9 CAA (Lys, Met, Thr, Trp, Val, Ile, Leu, His, and Phe) or only six CAA (Lys, Met, Thr, Trp, Val, and Ile) to meet the requirements. The CSC room was washed weekly, whereas the USC room had sow manure spread in the pens from the beginning of the study and was not washed throughout the experiment. Jejunum and colon digesta were sampled on day 21. Both jejunum and colon digesta were analyzed for ammonia nitrogen, short-chain fatty acids, and biogenic amines but only colon digesta was analyzed for microbiome composition (16s rRNA sequencing on MiSeq). Data were analyzed using R software for 16S rRNA and the MIXED procedure of SAS for microbial metabolites. Sanitation, CP content, and CAA supplementation patterns did not affect the diversity of colonic bacterial composition in weaned pigs. Pigs raised under USC had greater (P < 0.05) jejunal ammonia nitrogen concentration than those raised under CSC. Pigs fed LCP diets had reduced (P < 0.05) jejunal ammonia nitrogen concentration compared to those fed the HCP diet. Interactions between sanitation and dietary CP content were observed (P < 0.05) for: (1) jejunal acetate and (2) colonic spermidine and spermine, whereby (1) acetate concentrations decreased from NCP to LCP in pigs raised under the CSC but those concentrations increased under the USC, and (2) spermidine and spermine concentrations increased in LCP diets compared to HCP diet under USC, unlike CSC which did not show any difference between HCP and LCP. In conclusion, reducing dietary CP lowered ammonia nitrogen content regardless of sanitation and increased microbial metabolites in weaned pigs raised under USC. However, LCP diets with different CAA supplementation patterns did not affect bacterial diversity in weaned pigs, regardless of the hygienic conditions where the animals were housed.

Lowering dietary crude protein concentration by 3% to 4% units has been used as one of the strategies to promote growth and improve the gut health of weaned pigs. Undigested and endogenous protein could be available for microbial fermentation, and protein fermentation is considered detrimental to the gut health of the host animal. The unclean sanitary condition model mimics commercial raising conditions and stimulates a low-grade inflammatory and immune response. Ammonia nitrogen is one of the harmful metabolites derived by protein fermentation and pigs fed low-protein diets had decreased ammonia nitrogen than those fed high-protein diets. Also, pigs raised under unclean sanitation had greater ammonia nitrogen than those raised under clean sanitation. However, sanitation, protein content, and crystalline amino acids supplementation patterns did not affect the diversity of colonic bacterial composition in weaned pigs. The results obtained from the present study showed that a low protein diet could be used to improve gut health in weaned pigs.

Comparative effects of dietary methionine and cysteine supplementation on redox status and intestinal integrity in immunologically challenged-weaned pigs.

Sulfur-containing amino acids such as methionine and cysteine play critical roles in immune system and redox status. A body of evidence shows that metabolic aspects of supplemented Met and Cys may differ in the body. Therefore, the study aimed to investigate the effects of dietary Met and Cys supplementation in immunologically challenged weaned pigs. Forty weaned piglets (6.5 ± 0.3 kg) were randomly allocated to five treatment groups. The treatment included: (1) sham-challenged control (SCC), (2) challenged control (CC), (3) MET (CC + 0.1% DL-Met), (4) CYS (CC + 0.1% L-Cys), and (5) MET + CYS (CC + 0.1% DL-Met + 0.1% L-Cys). On day 7, all pigs were intramuscularly injected with either Escherichia coli O55:B5 lipopolysaccharides (LPS) or phosphate-buffered saline. Blood, liver, and jejunum samples were analyzed for immune response and redox status. The CC group had lower (P < 0.05) villus surface area and higher (P < 0.05) flux of 4-kDa fluorescein isothiocyanate dextran (FD4) than the SCC group. A lower (P < 0.05) glutathione (GSH) concentration was observed in the jejunum of pigs in the CC group than those in the SCC group. Dietary Cys supplementation increased (P < 0.05) villus surface area, GSH levels, and reduced (P < 0.05) the flux of FD4 in the jejunum of LPS-challenged pigs. Dietary Met supplementation enhanced (P < 0.05) hepatic GSH content. Pigs challenged with LPS in the MET group had lower serum IL-8 concentration than those in the CC group. There was a Met × Cys interaction (P < 0.05) in serum IL-4 and IL-8 concentrations, and Trolox equivalent antioxidant capacity. Dietary L-Cys supplementation restored intestinal integrity and GSH levels that were damaged by lipopolysaccharides administration. Dietary DL-Met supplementation improved hepatic GSH and reduced systemic inflammatory response, but antagonistic interaction with dietary L-Cys supplementation was observed in the inflammatory response and redox status.

Effects of dietary protein content and crystalline amino acid supplementation patterns on growth performance, intestinal histomorphology, and immune response in weaned pigs raised under different sanitary conditions.

The aim of this experiment was to investigate the effects of dietary crude protein (CP) contents and crystalline amino acids (CAA) supplementation patterns on growth performance, intestinal histomorphology, and immune response in weaned pigs under clean (CSC) or unclean sanitary conditions (USC). A total of 144 weaned pigs (6.35 ± 0.63 kg body weight) were assigned to 6 treatments in a 3 × 2 factorial arrangement based on CP content and sanitary conditions using a randomized complete block design, giving 8 replicates per treatment with 3 pigs per pen. Pigs were fed one of three diets for 21 d: one high CP (HCP; 22%) and two low CP (LCP; 19%) diets supplemented with 9 indispensable AA or only 6 AA (Lys, Met, Thr, Trp, Val, and Ile) as CAA. The CSC room was washed weekly, whereas the USC room had sow manure spread in the pens and was not washed throughout the experiment. Body weight and feed disappearance were recorded weekly. Blood was sampled from 1 pig per pen weekly, and the same pig was euthanized for jejunal tissues sampling on day 21. Pigs raised under USC had reduced (P < 0.05) average daily gain (ADG) and gain to feed ratio (G:F) in week 2, but contrary results that greater (P < 0.05) ADG and G:F were found in pigs under USC in week 3. Overall, there was an interaction where G:F did not differ between HCP and LCP under CSC, however, LCP decreased (P < 0.05) G:F compared to HCP under USC. Pigs fed the HCP diet had higher (P < 0.05) fecal scores than those fed the LCP diets throughout the experiment. Pigs fed the LCP had higher (P < 0.05) villus height to crypt depth ratio than those fed the HCP. An interaction was observed where goblet cell density in the jejunum was higher (P < 0.05) in pigs fed LCP than HCP under CSC, but no difference was found between HCP and LCP under USC. Different CAA supplementation patterns did not influence both growth performance and histomorphology. Pigs raised under USC had greater (P < 0.05) plasma interleukin (IL)-10 and IL-6 concentrations and reduced (P < 0.05) plasma tumor necrosis factor-alpha concentration. Also, the LCP diets resulted in a greater (P < 0.05) plasma IL-10 concentration. In conclusion, overall growth performance did not differ between HCP and LCP under CSC, but LCP diets reduced G:F under USC. Feeding LCP diets to weaned pigs improved gut morphology under USC and ameliorated systemic inflammation induced by USC, whereas CAA supplementation patterns did not affect growth performance and gut morphology.

Reducing dietary crude protein (CP) content reduces nitrogen excretion to the environment and undigested protein availability to enteric pathogens in the hind gut resulting in reduced postweaning diarrhea and improved gut health. To meet the amino acid requirements in low CP diets, supplementation of essential amino acids in nursery diets is necessary, and the availability of more feed-grade crystalline amino acids allows low CP diets to be practically formulated. The sanitation challenge model mimics commercial raising conditions and stimulates a low-grade inflammatory and immune response. Although low CP diets decreased the feed efficiency in pigs raised under unclean conditions in this study, improved gut histomorphology and ameliorated inflammatory response were obtained in pigs fed low CP diets. However, no effect was found in crystalline amino acids supplementation patterns. The outcomes of this study confirmed that low CP diets could be used to improve gut health and ameliorate immune response in weaned pigs.

Heat processing increased the digestibility of phosphorus in soybean expeller, canola meal, and canola expeller fed to growing pigs.

It is hypothesized that heat processing may increase P digestibility in different protein sources fed to growing pigs. A study was conducted to determine the apparent total tract digestibility (ATTD) and standardized total tract digestibility (STTD) of P in soybean expeller (SBE) produced from oil extraction using dry extrusion and expelling and to investigate the effects of heat treatment on the ATTD and STTD of P in SBE, canola meal (CM), and canola expeller (CE) fed to growing pigs. Thirty-six growing barrows with an initial body weight of 19.0 ± 1.0 kg (mean ± SD) were assigned to 1 of 6 experimental diets in a completely randomized design to give 6 replicates per diet. The experimental design was a 3 × 2 factorial arrangement including three oilseed meals with or without heat treatment. The diets were formulated to contain non-autoclaved or autoclaved (at 121 °C for 60 min) SBE, CM, and CE as the sole source of P. Limestone was included in diets to maintain a Ca:total P ratio of 1.3:1 across diets. Pigs were individually housed in metabolism crates for 12 d, including 7 d for adaptation and 5 d for total collection of feces. Pigs were offered their daily ration at 2.8 times their maintenance energy requirement. Data were analyzed using the PROC MIXED of SAS. Heat treatment increased (P < 0.05) the ATTD and STTD of P. Pigs fed the SBE diets had greater (P < 0.05) ATTD and STTD of P than pigs fed CM and CE diets. For the autoclaved ingredients, the values of STTD of P were 49.4%, 23.2%, and 25.8% for SBE, CM, and CE, respectively, whereas STTD of P in non-autoclaved SBE, CM, and CE were 48.5%, 20.2%, and 22.5%. Heat treatment increased (P < 0.05) the ATTD of Ca. In conclusion, heat treatment increased ATTD and STTD of P and ATTD of Ca in SBE, CM, and CE fed to growing pigs. The ATTD and STTD of P in SBE determined in the current study were 41.0% and 48.5%, respectively.

Diet complexity and l-threonine supplementation: effects on growth performance, immune response, intestinal barrier function, and microbial metabolites in nursery pigs.

The aim of this study was to investigate the effects of diet complexity and l-Thr supplementation level on the growth performance, immune response, intestinal barrier function, and microbial metabolites in nursery pigs. Thirty-two weaned pigs (body weight 7.23 ± 0.48 kg) were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement based on diet complexity (complex or simple) and dietary Thr content. The complex diet contained fish meal, plasma protein, and dried whey to mimic a conventional nursery diet. The simple diet was formulated with corn, wheat, and soybean meal and did not contain any animal products. l-Thr was supplemented to each diet to supply either 100% (STD Thr) or 115% (SUP Thr) of the NRC (2012) requirement for standardized ileal digestible Thr. Pigs were individually housed and fed experimental diets ad libitum for 14 d. Diet complexity, dietary Thr content, and their interactions were considered the main effects. Pigs fed the simple diet had greater (P < 0.05) plasma interleukin (IL)-10 and IL-6 concentrations compared with those fed the complex diet on days 7 and 14, respectively. Simple diet-fed pigs tended to show greater (P < 0.10) expression of genes encoding for tumor necrosis factor-α, claudin-1, and zonula occludens-1 in the jejunum compared with complex diet-fed pigs. The simple diet-fed pigs had greater (P < 0.05) concentrations of NH3-N in the jejunum digesta than did complex diet-fed pigs. The SUP Thr increased (P < 0.05) villus height and goblet cell (GC) density in villi and crypts in the jejunum and deepened (P < 0.05) crypts in the proximal colon. The SUP Thr resulted in the upregulation (P < 0.05) of occludin gene expression and a tendency toward the downregulation (P = 0.10) of IL-6 gene expression in the jejunum. Interactions (P < 0.05) between diet complexity and l-Thr supplementation level were observed in GC density in the crypt, NH3-N concentration in the jejunum, and the contents of acetate, propionate, and total volatile fatty acids in the colon. In conclusion, feeding a simple diet to nursery pigs resulted in systemic and intestinal inflammation. The SUP Thr diet did not normalize the simple diet-induced inflammation but improved gut integrity. SUP Thr seems to have greater benefits with a simple diet than with a complex diet. Therefore, SUP Thr in a simple diet could be a beneficial nutritional strategy for enhancing gut health.

Diet complexity and l-threonine supplementation: effects on nutrient digestibility, nitrogen and energy balance, and body composition in nursery pigs.

This study was conducted to investigate the effects of dietary complexity and l-Thr supplementation on energy and nutrient utilization in nursery pigs. Thirty-two nursery pigs (7.23 ± 0.48 kg) were randomly assigned to a 2 × 2 factorial treatment arrangement based on diet complexity (complex vs. simple) with different levels of l-Thr supplementation. The complex diet contained animal protein sources (e.g., fish meal and plasma) and a dairy product (e.g., dried whey) to mimic a conventional nursery diet. The simple diet was formulated with corn, wheat, and soybean meal. Both diets were supplemented with l-Thr to contain either 100% or 115% (SUP Thr) of the estimated standardized ileal digestible Thr requirement for 9 kg body weight pigs (NRC, 2012). The pigs were individually housed in metabolism crates and fed an experimental diet ad libitum for a 7-d adaptation period and 5 d of total but separate urine and fecal collection. On day 14, all pigs were euthanized to determine body composition. The diet complexity, l-Thr supplementation, and their interactions were considered main effects. Pigs fed the complex diet tended to exhibit greater (P < 0.10) apparent total tract digestibility (ATTD) of ash and urinary energy output than those fed the simple diet. The complex diet had greater (P < 0.05) digestible energy and net energy contents than the simple diet. Furthermore, the complex diet-fed pigs had lower (P < 0.05) plasma urea nitrogen concentration on day 14 than simple diet-fed pigs. The SUP Thr decreased (P < 0.05) ATTD of acid detergent fiber but trended (P < 0.10) toward a decrease in urinary nitrogen (N) output and an increase in N retention and body N mass. In conclusion, the simple diet for nursery pigs had lower digestible and net energy contents than a complex diet. The SUP Thr can improve N utilization and body protein deposition, irrespective of diet complexity.

Effect of dietary supplementation of xylanase in a wheat-based diet containing canola meal on growth performance, nutrient digestibility, organ weight, and short-chain fatty acid concentration in digesta when fed to weaned pigs.

This study was designed to determine the effect of dietary supplementation of xylanase on growth performance, nutrient digestibility, organ weight, digesta pH, and concentration of short-chain fatty acids (SCFA) of weaned pigs fed wheat-canola meal (CM) diets over a 35-d period. A total of 144 piglets (72 barrows and 72 gilts) weaned at 18 ± 2 d of age, with initial body weight (BW) of 6.2 ± 0.7 kg, received one of eight dietary treatments based on randomized complete block design. BW and feed intake were recorded weekly to calculate average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F). Treatments consisted of a control wheat-soybean meal-based diet and wheat-regular (RCM), dehulled (DCM), or coarse CM (CCM) without and with 16,000 BXU/kg xylanase (Econase XT). All diets contained 500 FTU/kg of phytase (Quantum Blue 5G) and titanium dioxide (0.3%). Apparent total tract digestibility (ATTD) of neutral detergent fiber (NDF), crude protein (CP), phosphorous (P), calcium (Ca), dry matter, and diet were determined. On day 35, one pig per pen was euthanized to evaluate the main factors of protein, xylanase supplementation, and sex on organ weight, ileal and colon digesta pH, and colon digesta concentrations of SCFA. The main factors did not affect growth performance. Xylanase supplementation improved nutrient digestibilities in all diets and increased ileal and colonic digesta pH without affecting the growth performance of weaned pigs fed wheat and CM-based diets. A protein-xylanase effect (P < 0.05) resulted in increasing the ATTD of NDF from 28% to 32% and from 29% to 37% for RCM and DCM, respectively. The ATTD of CP was greater (P < 0.05) with xylanase supplementation (75% vs. 70%). Xylanase supplementation increased ATTD of P and Ca. A three-way interaction (P < 0.05) for protein-xylanase-gender for colon pH, acetic, and propionic acid in the colon digesta of pigs indicated that, in addition to the protein source, piglet sex could have influenced how xylanase works. Xylanase supplementation increased (P < 0.05) the weight of the liver and spleen and tended (P < 0.10) to increase the size of the kidney. In conclusion, dietary supplementation of xylanase increased nutrient digestibility and digesta pH but did not influence the growth performance of weaned pigs fed wheat and CM-based diets over a 35-d period.

Effects of thermally oxidized canola oil and tannic acid supplementation on nutrient digestibility and microbial metabolites in finishing pigs1.

The present study investigated the effects of oxidized canola oil and tannic acid (TA) supplementation on nutrient digestibility and microbial metabolites in finishing pigs. Four experimental diets were formulated to include 5% of either fresh canola oil (0 mEq/kg of peroxide value) or oxidized canola oil (180 mEq/kg of peroxide value). An equal portion of each diet was supplemented with 0.1% TA. Eight pigs (initial body weight of 82.89 ± 2.26 kg) were surgically fitted with a T-cannula at the distal ileum. At the beginning of the experiments, all pigs were fed a nitrogen-free diet for 7 d. Then, they were assigned to 4 dietary treatments according to an incomplete replicated 4 × 3 Latin square design with 3 periods to give 6 replicates per treatment. Each experimental period lasted for 9 d, starting with a 5-d adaptation period followed by 2 consecutive 2-d periods for fecal (day 6 and day 7) and ileal digesta (day 8 and day 9) collection. The inclusion of oxidized oil reduced (P < 0.05) the standardized ileal digestibility (SID) of Pro and the apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of NDF. Also, pigs fed the oxidized oil tended to have lower (P ≤ 0.10) lactate and ammonia-nitrogen concentrations in the ileum and lower (P ≤ 0.10) acetate and propionate concentrations in the feces than those fed the fresh oil. The supplementation of 0.1% TA reduced (P < 0.05) the AID of NDF and the ATTD of gross energy, crude protein, and NDF. Additionally, the TA-supplemented diet tended to have a lower (P ≤ 0.10) digestible energy content than the nonsupplemented diet. Pigs fed the TA-supplemented diet showed reduced (P < 0.05) AID and SID of His, Cys, and Pro compared with those fed the nonsupplemented diet. There were interactive effects (P < 0.05) of oxidized oil and TA supplementation on acetate, isovalerate, total short chain fatty acids, and pH level in the ileum. In conclusion, the results indicated that the effects of oxidized oil are greater on microbial activities rather than nutrient digestibility. Although negative effects of dietary TA supplementation were observed in relation to nutrient digestibility, TA supplementation was found to ameliorate these negative effects of oxidized oil on microbial metabolites. Therefore, further studies are required to investigate the interaction between dietary oxidized oil and TA in relation with the gut microbiota in pigs.

Phosphatidylcholine could protect the defect of zearalenone exposure on follicular development and oocyte maturation.

Zearalenone (ZEA) is a well-known exogenous endocrine disruptor and can lead to severe negative effects on the human and animal reproductive process. Using a follicle culture model, we have previously shown that ZEA exposure significantly affected the follicular development and antrum formation but the underlying mechanisms are not well known. Therefore, in this study, we explored the metabolomic changes of granulosa cell (GC) culture media with or without ZEA exposure. The results showed that ZEA significantly increased phosphatidylcholine or phosphatidyl ethanolamine adducts in culture medium. A comprehensive analysis with the metabolome data from follicular fluid of small and large antral follicles showed that lyso phosphatidylcholine (LPC) was accumulated during follicle growth, but was depleted by ZEA exposure. Exogenous supplement with LPC to the follicle growth media or oocyte maturation media can partly protect the defect of ZEA exposure on follicular antrum formation and oocyte maturation. Taken together, our results demonstrate that ZEA exposure hinders the follicular growth and exogenous LPC can practically protect the defect of ZEA on follicular development and oocyte maturation.

Effects of levan-type fructan on growth performance, nutrient digestibility, diarrhoea scores, faecal shedding of total lactic acid bacteria and coliform bacteria, and faecal gas emission in weaning pigs.

BACKGROUND: The use of antibiotics as growth promoters in feed has been fully or partially banned in several countries. The objective of this study was to evaluate effects of levan-type fructan on growth performance, nutrient digestibility, faecal shedding of lactic acid bacteria and coliform bacteria, diarrhoea scores, and faecal gas emission in weaning pigs. A total of 144 weaning pigs [(Yorkshire × Landrace) × Duroc] were randomly allocated to four diets: corn-soybean meal-based diets supplemented with 0, 0.1, 0.5, or 1.0 g kg-1 levan-type fructan during this 42-day experiment. RESULTS: During days 0 to 21 and 0 to 42, average daily gain and average daily feed intake were linearly increased (P < 0.01) with increasing dietary levan-type fructan inclusion. The apparent total tract digestibility of dry matter, crude protein, and gross energy were linearly increased (P < 0.001) with increasing dietary levan-type fructan content. With increasing levels of levan-type fructan, faecal lactic acid bacteria counts were linearly increased (P = 0.001). CONCLUSION: The results indicate that dietary supplementation with increasing levan-type fructan enhanced growth performance, improved nutrient digestibility, and increased faecal lactic acid bacteria counts in weaning pigs linearly. © 2017 Society of Chemical Industry.

Therapeutic effects of glutamic acid in piglets challenged with deoxynivalenol.

The mycotoxin deoxynivalenol (DON), one of the most common food contaminants, primarily targets the gastrointestinal tract to affect animal and human health. This study was conducted to examine the protective function of glutamic acid on intestinal injury and oxidative stress caused by DON in piglets. Twenty-eight piglets were assigned randomly into 4 dietary treatments (7 pigs/treatment): 1) uncontaminated control diet (NC), 2) NC+DON at 4 mg/kg (DON), 3) NC+2% glutamic acid (GLU), and 4) NC+2% glutamic acid + DON at 4 mg/kg (DG). At day 15, 30 and 37, blood samples were collected to determine serum concentrations of CAT (catalase), T-AOC (total antioxidant capacity), H2O2 (hydrogen peroxide), NO (nitric oxide), MDA (maleic dialdehyde), DAO (diamine oxidase) and D-lactate. Intestinal morphology, and the activation of Akt/mTOR/4EBP1 signal pathway, as well as the concentrations of H2O2, MDA, and DAO in kidney, liver and small intestine, were analyzed at day 37. Results showed that DON significantly (P<0.05) induced oxidative stress in piglets, while this stress was remarkably reduced with glutamic acid supplementation according to the change of oxidative parameters in blood and tissues. Meanwhile, DON caused obvious intestinal injury from microscopic observations and permeability indicators, which was alleviated by glutamic acid supplementation. Moreover, the inhibition of DON on Akt/mTOR/4EBP1 signal pathway was reduced by glutamic acid supplementation. Collectively, these data suggest that glutamic acid may be a useful nutritional regulator for DON-induced damage manifested as oxidative stress, intestinal injury and signaling inhibition.

Effect of dietary crude protein level on jejunal brush border enzyme activities in weaned pigs.

Forty weaned pigs (7.0 ± 0.5 kg, mean ± SD) were used to determine the effects of feeding a low crude protein, amino acid-supplemented diet to piglets on the activities of jejunal brush border enzymes. Pigs were randomly allotted to two diets: a 222 g crude protein (HCP) per kg diet, or a 173 g crude protein per kg diet supplemented with amino acids (LCP). Pigs fed the HCP diet had higher small intestine weight compared with those fed the LCP diet on day 7 after weaning. Diet had no effect on the specific activities of jejunal sucrase, lactase, leucine aminopeptidase, aminopeptidase A, aminopeptidase N and dipeptidyl peptidase IV. The activities of sucrase and lactase decreased (p < 0.05) from day 3 after weaning to day 7, but the activities of leucine aminopeptidase and aminopeptidase N increased. The results showed that feeding a low protein diet supplemented with amino acids according to the ideal protein ratio to piglets had no negative effect on the development of jejunal brush border enzymes.