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BACKGROUND: Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E. coli (ETEC) F18 in a manner similar to carbadox. The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18. RESULTS: Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic. The relative abundance of metabolic markers of immune responses and nutrient metabolisms, such as amino acids and carbohydrates, were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups (q  2.0). In addition, pigs in antibiotic had a reduced (P

The objective of this study was to investigate the effects of dietary supplementation of Bacillus (B.) amyloliquefaciens on growth performance, diarrhea, systemic immunity, and intestinal microbiota of weaned pigs experimentally infected with F18 enterotoxigenic Escherichia coli (ETEC). A total of 50 weaned pigs (7.41 ± 1.35 kg BW) were individually housed and randomly allotted to one of the following five treatments: sham control (CON-), sham B. amyloliquefaciens (BAM-), challenged control (CON+), challenged B. amyloliquefaciens (BAM+), and challenged carbadox (AGP+). The experiment lasted 28 days, with 7 days of adaptation and 21 days after the first ETEC inoculation. ETEC challenge reduced (P < 0.05) average daily gain (ADG) of pigs. Compared with CON+, AGP+ enhanced (P < 0.05) ADG, while B. amyloliquefaciens supplementation tended (P < 0.10) to increase ADG in pigs from days 0 to 21 post-inoculation (PI). The ETEC challenge increased (P < 0.05) white blood cell (WBC) count on days 7 and 21 PI, while BAM+ pigs tended (P < 0.10) to have low WBC on day 7 PI and had lower (P < 0.05) WBC on day 21 PI compared with CON+. In comparison to AGP+ fecal microbiota, BAM+ had a lower (P < 0.05) relative abundance of Lachnospiraceae on day 0 and Clostridiaceae on day 21 PI, but a higher (P < 0.05) relative abundance of Enterobacyeriaceae on day 0. In ileal digesta, the Shannon index was higher (P < 0.05) in BAM+ than in AGP+. Bray-Curtis PCoA displayed a difference in bacterial community composition in ileal digesta collected from sham pigs vs. ETEC-infected pigs on day 21 PI. Pigs in BAM+ had a greater (P < 0.05) relative abundance of Firmicutes, but a lower (P < 0.05) relative abundance of Actinomycetota and Bacteroidota in ileal digesta than pigs in AGP+. Ileal digesta from AGP+ had a greater (P < 0.05) abundance of Clostridium sensu stricto 1 but lower (P < 0.05) Bifidobacterium than pigs in BAM+. In conclusion, supplementation of B. amyloliquefaciens tended to increase ADG and had limited effects on the diarrhea of ETEC-infected pigs. However, pigs fed with B. amyloliquefaciens exhibit milder systemic inflammation than controls. B. amyloliquefaciens differently modified the intestinal microbiota of weaned pigs compared with carbadox.

The objective of this study was to investigate supplementation of botanical blends (BB) comprised of 0.3% capsicum oleoresin and 12% garlic oil on gut microbiota and metabolomic profiles in serum and ileal mucosa of Escherichia coli infected pigs. Sixty weaned pigs were assigned to one of five treatments: negative control (CON-), positive control (CON+), dietary supplementation of 100 ppm BB1, 50 or 100 ppm BB2. All pigs, except CON-, were orally inoculated with 1010 CFU F18 ETEC/3-mL dose for 3 consecutive days after 7 d adaption. Feces, ileal digesta and cecal content were collected for 16S rRNA amplicon sequencing. Serum and ileal mucosa underwent primary metabolomics analysis. Supplementing 100 ppm BB1 increased (p < 0.05) relative abundances of Enterobacteriaceae and Escherichia-Shigella in ileum, and the relative abundances of Bacteroidota and Prevotellaceae in cecum than CON+ on d 5 post-inoculation (PI). Supplementing 100 ppm BB2 upregulated serum pinitol on d 4 PI and serum cholesterol and aminomalonic acids on d 21 PI, while supplementing 50 ppm BB2 reduced asparagine in ileal mucosa on d 5 PI than CON+. Supplementation with botanical blends modulated ileal and cecal microbiota and serum metabolomics profiles in weaned pigs under Escherichia coli challenge.

BackgroundThere 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.ResultsPigs in antibiotic carbadox or Coligo treatment groups had greater (P

BackgroundOur previous study has shown that supplementation of trace amounts of antibiotic exacerbated the detrimental effects of enterotoxigenic E. coli (ETEC) infection and delayed the recovery of pigs that may be associated with modified metabolites and metabolic pathways. Therefore, the objective of this study was to explore the impacts of trace levels of antibiotic (carbadox) on host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.ResultsThe multivariate analysis highlighted a distinct metabolomic profile of serum and colon digesta between trace amounts of antibiotic (TRA; 0.5 mg/kg carbadox) and label-recommended dose antibiotic (REC; 50 mg/kg carbadox) on d 5 post-inoculation (PI). The relative abundance of metabolomic markers of amino acids, carbohydrates, and purine metabolism were significantly differentiated between the TRA and REC groups (q

Our previous research reported that supplementation of Bacillus subtilis DSM 25841 promoted growth and disease resistance of weaned pigs under enterotoxigenic Escherichia coli (ETEC) challenge and its efficacy is comparable to carbadox. This follow-up study aimed to characterize the effects of ETEC infection, supplementing B. subtilis DSM 25841 or carbadox on intestinal microbiota of pigs. Forty-eight weaned pigs (6.17 ± 0.36 kg BW) were randomly allotted to one of four treatments: negative control (NC), positive control (PC), antibiotics (AGP, 50 mg/kg of carbadox), and direct fed microbials (DFM, 2.56 × 109  CFU/kg of B. subtilis). The experiment lasted 28 days with 7 days before and 21 days after first E. coli inoculation (day 0). Pigs in the PC, AGP, and DFM groups were orally inoculated with F18 ETEC for 3 consecutive days with 1010  CFU per dose per day. Fecal samples were collected on day -7, and day 7 and day 21 post inoculation, digesta samples were collected from jejunum, ileum, and distal colon on day 21 post inoculation to perform 16S rRNA sequencing. Sampling days and locations influenced (p < 0.05) Chao1 index and beta-diversity. Age increased (p  < 0.05) the relative abundance of Firmicutes but decreased (p < 0.05) the relative abundance of Bacteroidetes in feces. ETEC infection increased (p  < 0.05) the relative abundance of Proteobacteria in feces on day 7 post inoculation. AGP reduced (p < 0.05) relative abundance of Firmicutes and Lactobacillaceae in feces compared with PC and DFM. AGP reduced (p < 0.05) relative abundance of Bifidobacteriaceae in jejunum and ileum, while DFM reduced (p < 0.05) relative abundance of Actinomycetaceae in jejunum and Lachnospiraceae in ileum, compared with PC. Pigs fed with DFM had greater (p < 0.05) relative abundance of Ruminococcaceae, Veillonellaceae, Bifidobacteriaceae in jejunum, Lactobacillaceae in ileum and colon, and Bifidobacteriaceae in colon than pigs in AGP. Current results indicate that carbadox or B. subtilis had stronger influences on microbial diversity and composition in ileum than other intestinal segments and feces. Supplementation of B. subtilis could increase or maintain the relative abundance of beneficial bacteria in ileum compared with carbadox.

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) 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 (day 0). All pigs were orally inoculated with 1010 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 days -7, 0, 7, and 21 post first inoculation (PI), and ileal digesta and mucosal tissue collected on day 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

The experiment was conducted to investigate the effects of trace amounts of antibiotic on growth performance, diarrhea, systemic immunity, and intestinal health of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli. Weaned pigs (n = 34, 6.88 ± 1.03 kg body weight [BW]) were individually housed in disease containment rooms and randomly allotted to one of the three dietary treatments: nursery basal diet (CON) and two additional diets supplemented with 0.5 or 50 mg/kg carbadox to the nursery basal diet (TRA or REC), respectively. The experiment lasted 18 d with 7 d before and 11 d after the first E. coli inoculation. The E. coli F18 inoculum was orally provided to all pigs with a dose of 1010 colony-forming unit (CFU)/3 mL for three consecutive days. Fecal and blood samples were collected on day 0 before inoculation and days 2, 5, 8, and 11 postinoculation (PI) to test the percentage of β-hemolytic coliforms in total coliforms and complete blood cell count, respectively. Sixteen pigs were euthanized on day 5 PI, whereas the remaining pigs were euthanized at the end of the experiment to collect the jejunal and ileal mucosa and mesenteric lymph node for gene expression and bacterial translocation, respectively. Pigs in REC had greater (P < 0.05) final BW and lower (P < 0.05) overall frequency of diarrhea compared with pigs in the CON and TRA groups. Pigs in TRA had the lowest (P < 0.05) average daily gain and feed efficiency from day 0 to 5 PI, highest (P < 0.05) percentage of β-hemolytic coliforms in fecal samples on days 2 and 5 PI, and greatest (P < 0.05) bacterial colonies in mesenteric lymph nodes on day 11 PI compared with pigs in the CON and REC groups. Pigs in TRA had the greatest (P < 0.05) neutrophils on day 5 PI and higher (P < 0.05) white blood cell counts and lymphocytes than other groups on day 11 PI. Pigs in TRA had the greatest (P < 0.05) serum C-reactive protein on days 2 and 5 PI and serum tumor necrosis factor-α on day 5 PI, compared with pigs in the CON and REC groups. Pigs fed REC had increased (P < 0.05) mRNA expression of zona occludens-1 (ZO-1) and occludin (OCDN) and reduced (P < 0.05) interleukin-1 beta (IL1B), interleukin-6 (IL6), and tumor necrosis factor-alpha (TNFA) in ileal mucosa on day 5 PI, compared with the CON, whereas TRA upregulated (P < 0.05) mRNA expression of IL1B, IL6, and cyclooxygenase-2 (COX2) in the ileal mucosa on day 11 PI, compared with the REC. In conclusion, trace amounts of antibiotic may exacerbate the detrimental effects of E. coli infection on pig performance by increasing diarrhea and systemic inflammation of weanling pigs.

BackgroundPrevious research has shown that dietary supplementation of Bacillus spp. probiotics exerts beneficial effects on animals' growth. However, limited studies have evaluated the efficacy of Bacillus spp. on weaned pigs and their effects on host gut health and microbiome, and systemic immunity using a disease challenge model. The objective of this experiment was to investigate the effects of two Bacillus spp. strains (Bacillus subtilis DSM 32540 and Bacillus pumilus DSM 32539) on growth performance, diarrhea, intestinal health, microbiome, and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (ETEC).ResultsPigs in PRO1 (Bacillus subtilis DSM 32540) had greater (P

The study was conducted to investigate the efficacy of a probiotic Bacillus subtilis strain on growth performance, diarrhea, systemic immunity, and intestinal health of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli and to compare the efficacy of B. subtilis with that of carbadox. Weaned pigs (n = 48, 6.17 ± 0.36 kg body weight [BW]) were individually housed in disease containment rooms and randomly allotted to one of four dietary treatments: negative control (NC, control diet without E. coli challenge), positive control (PC, control diet with E. coli challenge), and supplementation of 50 mg/kg of carbadox (antibiotic growth promotor [AGP]) or 2.56 × 109 CFU/kg of B. subtilis probiotics (PRO). The experiment lasted for 28 d with 7 d before and 21 d after the first E. coli inoculation. Fecal and blood samples were collected on days 0, 3, 7, 14, and 21 post inoculation (PI) to analyze β-hemolytic coliforms and complete blood cell count, respectively. Diarrhea score was recorded daily for each pig to calculate the frequency of diarrhea. All pigs were euthanized at day 21 PI to collect jejunal and ileal mucosa for gene expression analysis. Pigs in AGP had greater (P < 0.05) BW on days 7, 14, and 21 PI than pigs in PC and PRO groups. Supplementation of PRO enhanced pigs' BW on day 21 PI compared with the PC. Escherichia coli F18 challenge reduced (P < 0.05) average daily gain (ADG) and feed efficiency from day 0 to 21 PI, while supplementation of carbadox or PRO enhanced ADG and feed efficiency in E. coli F18-challenged pigs from day 0 to 21 PI. Pigs in AGP and PRO groups had reduced (P < 0.05) frequency of diarrhea throughout the experiment and fecal β-hemolytic coliforms on day 7 PI than pigs in the PC. Pigs in PRO had greater (P < 0.05) gene expression of CLDN1 in jejunal mucosa than pigs in the PC. Supplementation of carbadox or PRO reduced (P < 0.05) the gene expression of IL6 and PTGS2 in ileal mucosa of E. coli-infected pigs compared with pigs in the PC. Pigs in the PRO group had lower (P < 0.05) white blood cell number and neutrophil count, and serum haptoglobin concentration on day 7 PI, and less (P < 0.05) monocyte count on day 14 PI, compared with PC. In conclusion, supplementation of probiotic B. subtilis could enhance disease resistance and promote the growth performance of weaned pigs under disease challenge conditions. The potential mechanisms include but not limited to enhanced gut barrier integrity and local and systemic immune responses of weaned pigs.

This experiment aimed to evaluate meat quality, fatty acid profile in back-fat, and fecal microbiota of growing-finishing pigs fed with liquid enzymatically digested food waste. Fifty-six crossbred pigs (approximately 32.99 kg body weight) were assigned to one of two treatments with seven replicate pens and four pigs per pen. Pigs were fed with control (corn-soybean meal diets) or food waste from d 0 to 53, while all pigs were fed with the control diet from d 53 to 79. The 16S rRNA sequencing was used to analyze microbiota of feces collected on d 0, 28, 53, and 79. Meat quality and carcass characteristics were measured in one pig per pen at the end of the experiment. Pigs fed with food waste contained more (p < 0.05) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in back-fat. Feeding food waste increased (p < 0.05) the relative abundances of Lachnospiraceae and Ruminococcaceae, but decreased (p < 0.05) the relative abundances of Streptococcaceae and Clostridiaceae in feces on d 29 or d 53. In conclusion, feeding enzymatically digested food waste did not affect pork quality, but provided more beneficial fatty acids to pork consumers and altered the fecal microbiota in growing-finishing pigs.

Oligosaccharides have been reported to possess ETEC receptor activity for bacterial adhesions; thus, additional supplementation may efficiently prevent enterotoxin-induced secretory diarrhea. Moreover, grafted polymers that combine multiple substances have been proposed for their potential synergistic effects on preventing human and animal diseases. Therefore, the objective of this experiment was to investigate dietary supplementation of oligosaccharide-based polymer on growth performance, diarrhea severity, intestinal health, and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (ETEC) when compared with in-feed antibiotics. Forty-eight pigs (7.23 ± 1.11 kg BW) were individually housed in disease containment rooms and randomly allotted to one of four treatments with 12 replicate pigs per treatment. The four dietary treatments were a nursery basal diet (CON), and 3 additional diets supplemented with 50 mg/kg antibiotics (Carbadox; CAR), 10 or 20 mg/kg of oligosaccharide-based polymer (OBP). The experiment lasted 18 d [7 d before and 11 d after the first inoculation (d 0)]. All pigs were orally inoculated with 3 mL of F18 ETEC for 3 consecutive days from d 0 post-inoculation (PI). Growth performance and diarrhea severity were measured throughout the experiment. Blood, lymph nodes, spleen, and intestinal mucosa samples were collected during the peak infection period (d 2 and d 5 PI) and recovery period (d 11 PI). Pigs in CAR or OBP treatment groups had greater (P < 0.05) body weight on d 5 or d 11 PI than pigs in the CON group, respectively. Supplementation of CAR or OBP 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 CON group. Pigs in CAR or OBP groups had reduced (P < 0.05) neutrophil counts and serum haptoglobin concentration compared to pigs in the CON group on d 2 and 5 PI. Pigs in OBP reduced (P < 0.05) total coliforms in mesenteric lymph nodes on d 5 and 11 PI, whereas pigs in CAR or OBP groups had reduced (P < 0.05) total coliforms in the spleen on d 11 PI compared with pigs in the CON group. On d 5 PI, pigs in the OBP 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 CON group. On d 11 PI, supplementation of CAR or OBP up-regulated (P < 0.05) gene expression of CLDN1 in jejunal mucosa, but OBP reduced (P < 0.05) IL6 gene expression in ileal mucosa compared to pigs in the CON group. In conclusion, supplementation of oligosaccharide-based polymer 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. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the dietary effects of Bacillus subtilis and antibiotics in intestinal microbiota of pigs experimentally infected with F18 E. coli. Forty-eight weaned pigs (6.17 ± 0.36 kg BW) were individually housed and randomly allotted in one of four treatments with 12 replicates per treatment: negative control (NC), positive control (PC), antibiotics, and B. subtilis (probiotics). Pigs in NC and PC were fed with basal diet without or with E. coli, respectively. Pigs with antibiotics and probiotics were challenged with E. coli and supplemented with 50 mg/kg of carbadox or 500 mg/kg of B. subtilis, respectively. After 7 days habituation period, pigs were inoculated with F18 E. coli at 1010 CFU/3 mL dose for three consecutive days. All pigs were euthanized to collect feces and digesta from jejunum, ileum, and colon on d 21 post-infection to perform 16S rRNA sequencing at the V4 hypervariable region. Downstream analysis was performed using QIIME2 (2019.4) and R. Kruskal-Wallis test followed by Conover test was used to analyze data in R. Colon digesta and feces have greater (P < 0.05) alpha diversity than ileal and jejunal digesta. No difference was observed among treatments at different intestinal sites. Bray-Curtis PCoA plots displayed pronounced clusters of all treatment groups throughout all sites. Bacteroidetes and Proteobacteria were more (P < 0.05) abundant but Firmicutes were less (P < 0.05) abundant in ileal digesta of pigs fed with antibiotics than pigs in probiotics. Firmicutes were more (P < 0.05) abundant in colon and feces of NC than of antibiotics. Bifidobacterium was least (P < 0.05) abundant throughout all sites and Prevotella 1 was most (P < 0.05) abundant in colon of pigs fed with antibiotics compared with other treatments. In conclusion, both B. subtilis and carbadox supplementation modified gut microbiota of weaned pigs challenged with F18 E. coli. However, the impacts are different and need further investigation. [ABSTRACT FROM AUTHOR]

The objective of this experiment was to investigate the effects of dietary supplementation of Bacillus amyloquefaciens on total and differential blood cell count in weaned pigs experimentally infected with a pathogenic E. coli. A total of 50 weaned pigs (7.41 ± 1.35 kg) were individually housed in disease containment rooms and randomly assigned to one of the 5 treatments: sham control (CON-), sham B. amyloquefaciens (BAM-), challenged control (CON+), challenged B. amyloquefaciens (BAM+) and challenged carbadox (CAR+). The experiment lasted 28 days with 7 days' adaptation and 21 days after the first E. coli inoculation. The doses of F18 E. coli inoculum were 1010 CFU/3 mL oral dose daily for 3 consecutive days. Whole blood samples were collected from all pigs on d -7, and d 0, 7, 14, and 21 post infection (PI) to measure total and differential blood cell count by complete blood count (CBC) analysis. Supplementation of BAM or CAR increased (P < 0.05) either the percentage or the number of lymphocytes on d 0 before E. coli inoculation. E. coli challenge increased (P < 0.05) white blood cell (WBC) count on d 7 and 21 PI, while supplementation of BAM tended (P < 0.10) to have low WBC on d 7 PI and had lower (P < 0.05) WBC on d 21 PI compared with CON+. Pigs in BAM+ also had lower (P < 0.05) neutrophil count on d 14 PI, pigs fed with CAR had lower (P < 0.05) neutrophil count on d 14 and 21 PI, compared with pigs in CON+. No difference was observed in red blood cell profile among all treatments throughout the experiment. In conclusion, pigs fed with B. amyloquefaciens have similar systemic immune response to pigs in antibiotic group and have relatively lower systemic inflammation caused by E. coli compared with control group. [ABSTRACT FROM AUTHOR]

The objective of this experiment was to investigate the effects of dietary supplementation of Bacillus amyloquefaciens on diarrhea and growth performance of weaned pigs experimentally infected with a pathogenic E. coli. A total of 50 weaned pigs (7.41 ± 1.35 kg) were individually housed in disease containment rooms and randomly assigned to one of the 5 treatments: sham control (CON-), sham B. amyloquefaciens (BAM-), challenged control (CON+), challenged B. amyloquefaciens (BAM+) and challenged carbadox (CAR+). The experiment lasted 28 days with 7 days' adaptation and 21 days after the first E. coli inoculation. The doses of F18 E. coli inoculum were 1010 CFU/3 mL oral dose daily for 3 consecutive days. Pigs were weighed on d -7, and d 0, 7, 14, and 21 PI and feed intake was recorded weekly to calculate growth performance. Diarrhea score was recorded twice daily ranging score from 1 (normal feces) to 5 (watery diarrhea). Fecal swabs were performed on each pig on d 2, 7, 14, and 21 PI to test the percentage of β-hemolytic coliforms in total coliforms. All data were analyzed with PROC MIXED of SAS. E. coli challenge reduced (P < 0.05) growth rate through the entire experiment. However, supplementation of CAR enhanced (P < 0.05) growth performance, while inclusion of BAM tended (P < 0.10) to increase growth performance of weaned pigs. No differences were observed in growth performance of pigs between BAM+ and CAR+ throughout the experiment. E. coli challenge did not change the percentage of β-hemolytic coliforms but increased (P < 0.05) frequency of diarrhea. However, no difference was observed in diarrhea among 3 dietary treatments. In conclusion, supplementing B. amyloquefaciens tended to enhance growth performance but had limited effects on diarrhea of weaned pigs challenged with E. coli. [ABSTRACT FROM AUTHOR]