Supplementation of oligosaccharide-based polymer enhanced growth and disease resistance of weaned pigs by modulating intestinal integrity and systemic immunity

Background There is a great demand for antibiotic alternatives to maintain animal health and productivity. The objective of this experiment was to determine the efficacy of dietary supplementation of a blood group A6 type 1 antigen oligosaccharides-based polymer (Coligo) on growth performance, diarrhea severity, intestinal health, and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (ETEC), when compared with antibiotics. Results Pigs in antibiotic carbadox or Coligo treatment groups had greater (P < 0.05) body weight on d 5 or d 11 post-inoculation (PI) than pigs in the control group, respectively. Supplementation of antibiotics or Coligo enhanced (P < 0.05) feed efficiency from d 0 to 5 PI and reduced (P < 0.05) frequency of diarrhea throughout the experiment, compared with pigs in the control group. Supplementation of antibiotics reduced (P < 0.05) fecal β-hemolytic coliforms on d 2, 5, and 8 PI. Pigs in antibiotics or Coligo groups had reduced (P < 0.05) neutrophil counts and serum haptoglobin concentration compared to pigs in the control group on d 2 and 5 PI. Pigs in Coligo had reduced (P < 0.05) total coliforms in mesenteric lymph nodes on d 5 and 11 PI, whereas pigs in antibiotics or Coligo groups had reduced (P < 0.05) total coliforms in spleen on d 11 PI compared with pigs in the control group. On d 5 PI, pigs in the Coligo group had greater (P < 0.05) gene expression of ZO1 in jejunal mucosa, but less (P < 0.05) mRNA expression of IL1B, IL6, and TNF in ileal mucosa, in comparison with pigs in the control group. Supplementation of antibiotics enhanced (P < 0.05) the gene expression of OCLN in jejunal mucosa but decreased (P < 0.05) IL1B and IL6 gene expression in ileal mucosa, compared with the control. On d 11 PI, supplementation of antibiotics or Coligo up-regulated (P < 0.05) gene expression of CLDN1 in jejunal mucosa, but Coligo reduced (P < 0.05) IL6 gene expression in ileal mucosa compared to pigs in the control group. Conclusions Supplementation of Coligo improved growth performance, alleviated diarrhea severity, and enhanced gut health in weaned pigs infected with ETEC F18 in a manner similar to in-feed antibiotics.

A Single Nucleotide Polymorphism in Promoter of Porcine ARHGAP24 Gene Regulates Aggressive Behavior of Weaned Pigs After Mixing by Affecting the Binding of Transcription Factor P53

Background: Weaned pigs often have more aggressive behavior after mixing, which has negative effects on animal welfare and growth performance. Identification of functional single nucleotide polymorphisms (SNPs) related to aggressive behavior of pigs would provide valuable molecular markers of aggressive behavioral trait for genetic improvement program. Rho GTPase Activating Protein 24 (ARHGAP24) gene plays an important role in regulating the process of axon guidance, which may impact aggressive behavior of pigs. Results: By re-sequencing the entire coding region, partially adjacent introns and the 5’ and 3’ flanking regions, 6 and 4 SNPs were identified in the 5’ flanking region and 5’ untranslated region (UTR) of porcine ARHGAP24 gene, respectively. Association analyses revealed that 9 SNPs were significantly associated with aggressive behavioral traits (P = < 1.00 × 10−4 - 4.51 × 10−2), and their haplotypes were significantly associated with aggressive behavior (P = < 1.00 × 10−4 - 2.99 × 10−2). The core promoter region of ARHGAP24 gene was identified between -670 bp and -1113 bp. Furthermore, the luciferase activity of allele A of rs335052970 was significantly less than that of allele G, suggesting the transcriptional activity of ARHGAP24 gene was inhibited by allele A of rs335052970. It was identified that the transcription factor p53 bound to the transcription factor binding sites (TFBSs) containing allele A of rs335052970. In porcine primary neural cells, p53 bind to the target promoter region of ARHGAP24 gene, reduce its promoter transcriptional activity, and then reduce its messenger RNA (mRNA) and protein expression through axon guidance pathway.Conclusion: The results demonstrated that ARHGAP24 gene had significant genetic effects on aggressive behavioral traits of pigs. Therefore, rs335052970 in ARHGAP24 gene can be used as a molecular marker to select less aggressive pigs and improve animal welfare.

Comparative proteomic study of pig muscle proteins during growth and development of an animal

The production of high-quality pork is closely related to the growth and development of muscle tissue. The present article provides a comparative proteomic research of l. dorsi, b. femoris, m. brachiocephalicus during the pigs’ growth and development (at age of 60 days and 180 days). This work was supported by data of electrophoretic methods: one-dimensional electrophoresis according to Laemmli with densitometric assessment in the ImageJ software and two-dimensional electrophoresis according to O’Farrell method with its further processing on the software ImageMaster. The mass spectrometric identification was conducted with the help of the high-performance liquid chromatography (HPLC) system connected to a mass spectrometer; further the data were interpreted by search algorithm Andromeda. When comparing frequency diagrams of one-dimensional electrophoregrams of all three muscle tissues of weaned pigs, the greatest difference was observed for the muscle sample l. dorsi. Comparison of diagrams of muscle tissue samples taken for mature pigs showed a great similarity of all three studied muscles samples. Within the framework of the research, the Fold indicator was calculated. The exceeding its value by more than 2 units is generally considered to be a statistically significant difference. When analyzing two-dimensional electrophoretograms of weaned pigs’ muscles, 18 protein fractions were revealed with Fold > 2. When examining the muscle tissue of mature pigs, 15 of those proteins were found; the differences were mostly detected in the minor protein fractions. The mass spectrometric analysis of the cut bands with well-pronounced differences from the onedimensional electrophoretogram revealed 214 proteins involved to a greater extent in cellular and metabolic processes, physical activity and localization. Growth and development protein — semaphorin‑6B (96.78 kDa) — was revealed in muscle tissue of l. dorsi, a. Also in l. dorsi and b. femoris the growth and development proteins were found: cadherin‑13 (78.23 kDa), cadherin‑7 (87.01 kDa), the F‑actin-cap protein beta subunit (30.66 kDa), and two uncharacterized proteins at 65.60 kDa and 63.88 kDa.

Effects of a blend of essential oils, medium-chain fatty acids and a toxin-adsorbing mineral on diarrhea and gut microbiome of weanling pigs experimentally infected with a pathogenic E. coli

A proprietary antimicrobial feed additive comprised of essential oils, medium-chain fatty acids, and a toxin-adsorbing mineral showed promising bacteriostatic and bactericidal effects in vitro. This study investigated the impacts of supplementing this blend on growth, gut microbiome, and enteric disease resilience in weaned pigs experimentally challenged with an enterotoxigenic Escherichia coli (ETEC). Thirty-six weanling pigs (6.88 ± 0.30 kg body weight (BW)) blocked by weight and gender were assigned to one of three dietary treatments: control or dietary supplementation with 0.25% or 0.50% of the antimicrobial blend. This study lasted 28 d with 7 d before and 21 d after the first ETEC inoculation (d 0). All pigs were orally inoculated with 10 10 cfu F18+ ETEC/3-mL dose for 3 consecutive days. Growth performance data and diarrhea scores were recorded throughout the experiment. Fecal samples collected on d -7, 0, 7 and 21 post first inoculation (PI), and ileal digesta and mucosal tissue collected on d 21 PI were further analyzed for gut microbiome using 16S rRNA sequencing. All data, except for frequency of diarrhea and gut microbiome, were analyzed by ANOVA using the PROC MIXED of SAS. The Chi-square test was used for analyzing frequency of diarrhea. Gut microbiome data were analyzed using QIIME2 and visualized using the R program. Dietary supplementation of 0.25% or 0.5% of the antimicrobial blend increased (P &lt; 0.05) feed efficiency on d 14 to 21 PI of ETEC and reduced (P &lt; 0.05) frequency of diarrhea during the study. Compared to the control group, adding 0.5% dietary antimicrobial blend increased (P &lt; 0.05) relative abundance of Firmicutes but reduced (P &lt; 0.05) Bacteroidetes and Proteobacteria in feces on d 7 PI. Pigs that received the antimicrobial blend also had higher (P &lt; 0.05) relative abundance of Lactobacillaceae, but lower (P &lt; 0.05) relative abundance of Enterobacteriaceae in feces on d 7 PI than pigs in control. In conclusion, supplementation of this antimicrobial blend at 0.5% reduced incidence of severe diarrhea in weaned pigs challenged with F18 ETEC and enhanced feed efficiency of weaned pigs at the last week of the experiment. Supplementation of this antimicrobial blend also modified the microbiota diversity in feces and ileal mucosa of weaned pigs.

Supplementation with Exogenous Catalase from Penicillium notatum in the Diet Ameliorates Lipopolysaccharide-Induced Intestinal Oxidative Damage through Affecting Intestinal Antioxidant Capacity and Microbiota in Weaned Pigs

Exogenous CAT derived from microorganisms has been widely used in food, medicine, and other industries. Recent study also found that exogenous CAT supplementation could improve growth performance and antioxidant capacity of weaned pigs.

Fermented Alfalfa Meal Instead of “Grain-Type” Feedstuffs in the Diet Improves Intestinal Health Related Indexes in Weaned Pigs

Corn and soybean meal are the two main components in formula feed of farm animals, leading to a serious food competition between humans and livestock. An alternative may be to encourage the utilization of unconventional feedstuff in animal diet. In the current study, we evaluated the utilization of fermented alfalfa meal (FAM) in weaned pigs. Twenty weaned piglets (separately caged) were randomly divided into two groups. Pigs in the control group (CON) were fed corn-soybean meal diet, and part of corn and soya protein concentrate in the diet of another group was replaced by 8% FAM. After 40 days of feeding, the average feed intake of FAM pigs was increased (P &gt; 0.05), and the villus height (VH) of jejunum and duodenum, crypt depth (CD), and VH/CD in FAM pigs was improved compared to the CON group (P &lt; 0.05). The increase (P &lt; 0.05) of goblet cells in the jejunum of FAM pigs was positively correlated with the expression of MUC-2 gene (R = 0.9150). The expression of genes related to immunity (IRAK4, NF-κB, and IL-10) and intestinal barrier (Occludin and MUC-2) in the jejunum, as well as the expression of ZO-1 and MUC-2 in the colon of these pigs, also showed increase (P &lt; 0.05) compared to CON pigs, which was accompanied by the decrease (P &lt; 0.05) of LPS concentration in the serum. The elevated proportion of CD3+ and CD8+ T-lymphocyte subsets in spleen (P &lt; 0.05) confirmed the improvement of systemic immune function in FAM pigs. In addition, FAM pigs have a higher β-diversity of microbial community (P &lt; 0.05) and promoted enrichment of probiotics such as Lactobacillus that positively was correlated with acetate concentration in the colon over CON pigs. In summary, partially replacement of expanded corn and soya protein concentrate with FAM (8%) may benefit the intestinal barrier and immune function of weaned pigs without affecting their growth. Our findings also provide evidence of the feasibility of FAM as a dietary component in pigs to reduce the consumption of grain.

Effect of dietary organic acids supplementation on growth performance, nutrient digestibility, and gut morphology in weaned pigs

This study was conducted to investigate the effects of dietary supplementation with different organic acid (<b>OA</b>) mixtures on growth performance, nutrient digestibility, and gut morphology in weaned pigs. A total of 56 weaned pigs (7.93 ± 1.04 kg body weight) were assigned to four dietary treatments with seven replicates per treatment for a 35-d study conducted over two phases; phase 1 (d 1 to 14) and phase 2 (d 14 to 35). Diets consisted of 1) a corn-soybean meal-basal diet without any additive (negative control, <b>NC</b>); 2) NC + formic and propionic acids (<b>TRT1</b>); 3) NC + butyric, formic, and propionic acids (<b>TRT2</b>); and 4) NC + antibiotic (positive control, <b>PC</b>). During the overall period, the gain to feed ratio of pigs fed the PC and TRT1 diets tended to be higher (<i>P</i> = 0.059) than that of the NC diet. However, OA supplementation had no effects on nutrient digestibility. Jejunal villus height to crypt depth ratio was higher (<i>P</i> < 0.05) in pigs fed the TRT1 diet than those fed the NC diet. In conclusion, dietary OA supplementation tended to increase overall feed efficiency and improved gut morphology in weaned pigs.

RESPONSE OF WEANED PIGS TO DIFFERENT DIETARY PROTEIN SOURCES AND DIET TYPES

Pig is one of the most consumed meat across the globe. However, its production is limited by high cost of feed ingredients. Hence, the search for alternative cheap feed ingredients without any loss to animal’s performance is on the increase. The influence of different protein sources and feed types (wet or dry) was investigated on the growth performance, haematology and serum biochemistry of weaned pigs. Weight gain and feed conversion ratio were higher for soybean and full-fat soya-based diets. Feed and water intake were not significantly different amongst treatments. Nature of feed did not influence weight gain, feed intake and feed conversion ratio. Groundnut cake meal may compare with soybean meal in pig’s diet, but it may have to be supplemented with lysine, methionine or fish meal.

Effects of Three-Layer Encapsulated Tea Tree Oil on Growth Performance, Antioxidant Capacity, and Intestinal Microbiota of Weaned Pigs

Tea tree oil (TTO) exerts key roles in improving growth performance of pigs. However, knowledge is limited regarding comparative effects of Encp TTO and Un-encp TTO supplementation on growth performance of pigs. A study determined the effects of TTO or its capsulation on growth performance, antioxidant capacity, and intestinal microbiome of weaned pigs. A total of 144 healthy pigs (8.5 ± 0.24 kg) were subjected to four treatments for a 28-d trial with six replicates per treatment and six pigs per pen: negative control, NC; positive control, PC (antibiotic supplemented); Un-encp TTO (supplemented with unencapsulated TTO); Encp TTO (supplemented with encapsulated TTO). NC, TTO, and PC treatments were compared with regard to improved average daily gain (ADG), average daily feed intake (ADFI), feed conversion rate, nutrient digestibility, and intestinal morphology (p &lt; 0.05) and decreased diarrhea rate. TTO- and PC-treated pigs had higher levels of serum superoxide dismutase, glutathione peroxidase, and immunoglobulin G; lower levels of liver aspartate aminotransferase and alanine aminotransferase; and improved concentrations of interleukin 10 (IL-10), tumor necrosis factor α, and IL-1β (p &lt; 0.05). TTO- and PC-treated pigs had higher abundance of beneficial bacterial species Subdoligranulum and lower abundance of diarrhea associated species Escherichia–Shigella in cecal and colonic digesta (p &lt; 0.05). Encapsulation of TTO preserved more activities of TTO than its unencapsulated counterpart by showing higher ADG, ADFI, and feed conversion rate during day 1 (d1) to d14 (p &lt; 0.05) and tended to lower diarrhea rate (p = 0.083) and improve villous height/crypt depth (VH/CD) ratio (p = 0.089) in jejunum. Encapsulation of TTO also improved antioxidant indexes and decreased liver injury and inflammation accordingly (p &lt; 0.05). Encapsulated TTO-treated pigs had higher abundance of beneficial Clostridium_sensu_stricto_1 and lower the abundance of harmful Escherichia–Shigella in cecal and colonic digesta (p &lt; 0.05). Our results demonstrated TTO benefits on improving growth performance of weaned pigs and further proved that encapsulation of TTO was superior to its unencapsulated counterpart at multiples. Encapsulated TTO was similar to the PC group and could be potentially an alternative of feed antibiotics for weaned pigs.