scholarly journals Trace Amounts of Antibiotic Altered Metabolomic and Microbial Profiles of Weaned Pigs Infected with a Pathogenic E. coli­­­­­­­

Author(s):  
Kwangwook Kim ◽  
Cynthia Jinno ◽  
Peng Ji ◽  
Yanhong Liu

Abstract Background: Our previous study has shown that supplementation of trace amounts of antibioticexacerbated 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. Results: The multivariate analysis highlighted a distinct metabolomic profile of serum and colon digesta between trace amounts of antibiotic (TRA) and label-recommended dose antibiotic (REC) 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 < 0.2). In addition, pigs in REC group had the highest (P < 0.05) relative abundance of Lactobacillaceae and tended to have increased (P < 0.10) relative abundance of Lachnospiraceae in the colon digesta on d 5 PI. On d 11 PI, pigs in REC had greater (P < 0.05) relative abundance of Clostridiaceae compared with other groups, whereas had reduced (P < 0.05) relative abundance of Prevotellaceae than pigs in control group.Conclusions: Trace amounts of antibiotic resulted in differential metabolites and metabolic pathways that may be associated with its slow responses against ETEC F18 infection. The altered gut microbiota profiles by label-recommended dose antibiotic may contribute to the promotion of disease resistance in weaned pigs.

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 183-183
Author(s):  
Cynthia Jinno ◽  
Yijie He ◽  
Xunde Li ◽  
Yanhong Liu

Abstract The objective of this study was to investigate the effects of supplementing Bacillus subtilis on fecal microbiota of pigs experimentally infected with F-18 Escherichia coli (E. coli), in comparison to carbadox. Forty-eight weaned pigs (6.17 ± 0.36 kg BW) were individually housed and randomly allotted to one of four treatment (n =12): negative control (NC), positive control (PC), antibiotics (50 mg/kg of carbadox), and direct fed microbials (DFM, 500 mf/kg of Bacillus subtilis). The experiment lasted 28 days with 7 days before and 21 days after first E. coli inoculation (d 0). Pigs in the NC, PC, and DFM groups were orally inoculated with F18 E. coli for 3 consecutive days with 1010 CFU/3 mL/dose. Fecal samples were collected on d -7 and 0 before E. coli inoculation, and d 7 and 21 post inoculation (PI). DNA were extracted from all fecal samples to perform 16S rRNA gene sequencing at the V4 hypervariable region. All data were analyzed with QIIME2 (2019.4) and R. Chao1 index was greatest (P &lt; 0.05) in feces collected on d 0 before E. coli inoculation and lowest (P &lt; 0.05) on d -7 feces. Pigs supplemented with DFM had lower (P &lt; 0.05) Chao1 index than pigs fed with antibiotics on d 21 PI. Bray-Curtis PCoA displayed separate clusters among days but overlaps among treatments. Bacteroidetes and Proteobacteria were most (P &lt; 0.05) abundant on d -7 and lowest (P &lt; 0.05) on d 21 PI. However, Actinobacteria and Firmicutes were most (P &lt; 0.05) abundant on d 21 PI. Pigs in the NC and DFM groups had greater (P &lt; 0.05) relative abundance of Firmicutes than pigs fed with antibiotics on d 0 and 7. Supplementation of antibiotics reduced (P &lt; 0.05) the relative abundance of Lactobacillaceae compared with other treatments on d 0 PI. In conclusion, both animal age and dietary treatments influenced the fecal microbiome of weaned pigs.


2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 286-286
Author(s):  
Kwangwook Kim ◽  
Sungbong Jang ◽  
Yanhong Liu

Abstract Our previous studies have shown that supplementation of low-dose antibiotic growth promoter (AGP) exacerbated growth performance and systemic inflammation of weaned pigs infected with pathogenic Escherichia coli (E. coli). The objective of this experiment, which is extension of our previous report, was to investigate the effect of low-dose AGP on gene expression in ileal mucosa of weaned pigs experimentally infected with F18 E. coli. Thirty-four pigs (6.88 ± 1.03 kg BW) were individually housed in disease containment rooms and randomly allotted to one of three treatments (9 to 13 pigs/treatment). The three dietary treatments were control diet (control), and 2 additional diets supplemented with 0.5 or 50 mg/kg of AGP (carbadox), respectively. The experiment lasted 18 d [7 d before and 11 d after first inoculation (d 0)]. The F18 E. coli inoculum was orally provided to all pigs with the dose of 1010 cfu/3 mL for 3 consecutive days. Total RNA [4 to 6 pigs/treatment on d 5; 5 to 7 pigs/treatment on 11 post-inoculation (PI)] was extracted from ileal mucosa to analyze gene expression profiles by Batch-Tag-Seq. The modulated differential gene expression were defined by 1.5-fold difference and a cutoff of P &lt; 0.05 using limma-voom package. All processed data were statistically analyzed and evaluated by PANTHER classification system to determine the biological process function of genes in these lists. Compared to control, supplementation of recommended-dose AGP down-regulated genes related to inflammatory responses on d 5 and 11 PI; whereas, feeding low-dose AGP up-regulated genes associated with negative regulation of metabolic process on d 5, but down-regulated the genes related to immune responses on d 11 PI. The present observations support adverse effects of low-dose AGP in our previous study, indicated by exacerbated the detrimental effects of E. coli infection on pigs’ growth rate, diarrhea and systemic inflammation.


2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 90-90
Author(s):  
Kwangwook Kim ◽  
Yijie He ◽  
Cynthia Jinno ◽  
Seijoo Yang ◽  
Xunde Li ◽  
...  

Abstract The objective of this experiment was to investigate dietary supplementation of oligosaccharide-based polymer on growth performance, diarrhea, and fecal β-hemolytic coliforms of weaned pigs experimentally infected with a pathogenic F18 Escherichia coli (E. coli). 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 (control), and 3 additional diets supplemented with 50 mg/kg Mecadox (AGP), 10 or 20 mg/kg of oligosaccharide-based polymer. The experiment lasted 18 d [7 d before and 11 d after the first inoculation (d 0)]. The doses of F18 E. coli inoculum were 1010 cfu/3 mL oral dose daily for 3 days. Growth performance was measured on d -7 to 0 before inoculation, and d 0 to 5 and 5 to 11 post-inoculation (PI). Diarrhea score (DS; 1, normal, to 5, watery diarrhea) was daily recorded for each pig. Fecal samples were collected on d 2, 5, 8, and 11 PI to test the percentage of β-hemolytic coliforms in total coliforms. All data were analyzed by ANOVA using the PROC MIXED of SAS with pig as the experimental unit. Inclusion of oligosaccharide-based polymer linearly increased (P &lt; 0.05) ADFI on d 0 to 5 PI, and feed efficiency on d 0 to 5 PI and d 5 to 11 PI (P = 0.07), compared with the control. Supplementation of AGP or oligosaccharide-based polymer reduced (P &lt; 0.01) frequency of diarrhea of pigs from d 0 to 11 PI. No differences were observed in overall growth performance and percentage of fecal β-hemolytic coliforms on d 8 PI among pigs in AGP and oligosaccharide-based polymer treatments. In conclusion, supplementation of oligosaccharide-based polymer enhanced feed efficiency and reduced diarrhea of weaned pigs infected with a pathogenic E. coli.


2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 168-169
Author(s):  
Cynthia Jinno ◽  
Braden T Wong ◽  
Martina Kluenemann ◽  
Xunde Li ◽  
Yanhong Liu

Abstract 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 &lt; 0.05) either the percentage or the number of lymphocytes on d 0 before E. coli inoculation. E. coli challenge increased (P &lt; 0.05) white blood cell (WBC) count on d 7 and 21 PI, while supplementation of BAM tended (P &lt; 0.10) to have low WBC on d 7 PI and had lower (P &lt; 0.05) WBC on d 21 PI compared with CON+. Pigs in BAM+ also had lower (P &lt; 0.05) neutrophil count on d 14 PI, pigs fed with CAR had lower (P &lt; 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.


2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Kwangwook Kim ◽  
Yijie He ◽  
Cynthia Jinno ◽  
Lauren Kovanda ◽  
Xunde Li ◽  
...  

Abstract 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.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 232-232
Author(s):  
Kwangwook Kim ◽  
Yijie He ◽  
Cynthia Jinno ◽  
Minho Song ◽  
Peng Ji ◽  
...  

Abstract Our previous studies have shown that exposure to low-dose antibiotic growth promoter (AGP) may exacerbate the negative effects of Escherichia coli (E. coli) infection on weaned pigs. The objective of this experiment was to investigate the effects of low-dose AGP on serum metabolomic profiles of weaned pigs experimentally infected with F18 E. coli. Thirty-four pigs (6.88 ± 1.03 kg BW) were individually housed in disease containment rooms and randomly allotted to one of three treatments with 9–13 replicate pigs per treatment. The three dietary treatments were control diet (control) and 2 additional diets supplemented with 0.5 or 50 mg/kg of AGP (carbadox), respectively. The experiment lasted 18 d [7 d before and 11 d after first inoculation (d 0)]. The F18 E. coli inoculum was orally provided to all pigs with the dose of 1010 cfu/3 mL for 3 consecutive days. Blood samples were collected on d 0 before E. coli inoculation and on d 5 and 11 post-inoculation (PI). Serum metabolomics were analyzed for untargeted metabolomics by gas chromatography-time of flight-mass spectrometer (GC-TOF-MS). All processed data were statistically analyzed and evaluated by online MetaboAnalyst tool. No significant differences were observed in serum metabolites between control and low-dose AGP throughout the experiment. Supplementation of high-dose AGP changed the concentrations of several serum metabolites (P &lt; 0.05; arabitol, guanine, and xylitol) compared with control pigs on d 5 PI. Further metabolic pathway enrichment analysis showed that low-dose AGP modified (P &lt; 0.05) pentose phosphate pathway, DNA synthesis in T and B lymphocytes, bile acid metabolism, and amino acid metabolism in E. coli infected pigs, compared with high-dose AGP. In conclusion, the modification of serum metabolites and metabolic pathways by low-dose AGP may be involved in reduced growth performance, exacerbated diarrhea and systemic inflammation of weaned pigs induced by E. coli infection.


Animals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1858
Author(s):  
Yibing Wang ◽  
Yang Wang ◽  
Xiajing Lin ◽  
Zhongyong Gou ◽  
Qiuli Fan ◽  
...  

This study was conducted to evaluate the effects of acidifier (benzoic acid, BA), amylase (AL) and their combination as substitutes for antibiotics on growth performance, antioxidation, nutrient digestion and gut microbiota of yellow-feathered broilers. A total of 1440 twenty-one-day-old broilers were randomly allocated to six treatments. Broilers in the control group (CON) were fed a basal diet, whereas birds in the other five groups were fed the basal diet supplemented with antibiotic (zinc bacitracin, AT, 40 mg/kg), BA (2000 mg/kg), low level AL (AL-L, 300 mg/kg), high level AL (AL-H, 500 mg/kg) and the combination of AL-H and BA (BA+AL-H). The experimental animals were killed at the end of the trial (21 day-63 day) then blood samples were collected from two birds per pen. Bird weight, feed intake and survival rate were recorded on pen basis. Growth performance was not significantly influenced by AT, BA, AL-L, AL-H or BA+AL-H. Plasma uric acid (UA) was decreased from CON by all treatments; the activity of AKP in plasma was also lowered by AT, BA, AL-H and BA+AL-H. Plasma activity of LDH was reduced by BA. In the jejunal mucosa, Na+K+-ATP activity was increased by BA, AL-L, AL-H and BA+AL-H. Mucosal activities of T-AOC and CAT were increased with AL-L and AT supplementation, respectively. Additionally, the relative abundance of Escherichia coli (E. coli) in the cecal contents was reduced by BA+AL-H and, with the exception of AL-H, all treatments increased the relative abundance of Lactobacillus. In conclusion, dietary AT, BA, AL-L, AL-H or BA+AL were effective in improving the antioxidant capacity, nutrient digestion and gut microbiota composition. No significant differences were observed in the tested variables between AT and other treatments, indicating that BA, AL and their combination may be alternatives to dietary inclusion of zinc bacitracin. Dietary addition of 500 mg/kg AL and 2000 mg/kg BA was an optimum supplementation dose.


2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 203-203
Author(s):  
Kwangwook Kim ◽  
Yanhong Liu

Abstract Our previous studies have shown that supplementation of Bacillus subtilis enhanced growth rate, improved gut barrier function, and modified colon microbiome of weaned pigs infected with pathogenic Escherichia coli (E. coli). The objective of this experiment was to investigate the effects of Bacillus subtilis on colon digesta metabolomic profiles of weaned pigs experimentally infected with F18 E. coli. Forty-eight pigs (6.73 ± 0.77 kg BW) were individually housed in disease containment rooms and randomly allotted to one of the four treatments (12 pigs/treatment). Four treatments included negative control (NC), positive control (PC), low-dose (1.28 × 109 CFU Bacillus subtilis/kg feed), and high-dose (2.56 × 109 CFU Bacillus subtilis/kg feed). The experiment lasted 18 d [7 d before and 11 d after first inoculation (d 0)]. The F18 E. coli inoculum was orally provided to all pigs with the dose of 1010 cfu/3 mL for 3 consecutive days, except NC. Twenty-four pigs (6 pigs/treatment) were euthanized on d 5 post-inoculation (PI) and the remained pigs were euthanized on d 11 PI to collect colon digesta for the analysis of metabolomic profiles by gas chromatography time of flight-mass spectrometer (GCTOF-MS). All processed data were statistically analyzed and evaluated by online MetaboAnalyst tool. No significant differences were observed in the metabolites between NC and PC on d 5 and 11 PI. Compared with PC, low- and high-dose Bacillus subtilis reduced (Fold change &gt; 1.5; FDR &lt; 0.20) four metabolites (proline, 2-hydroxyglutaric acid, lysine, and glutamic acid) and two metabolites (ribose, and D-xylulose) in colon digesta on d 5 PI, respectively. These metabolites were related to aminoacyl-tRNA-biosynthesis, arginine and proline metabolism, and lysine degradation. In conclusion, supplementation of Bacillus subtilis modified the levels of microbial metabolites associated with amino acid metabolism in colon digesta of pigs.


2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 74-75
Author(s):  
Yijie He ◽  
Kwangwook Kim ◽  
Cynthia Jinno ◽  
Zhaohai Wu ◽  
Rose Whelan ◽  
...  

Abstract The experiment was conducted to investigate the effects of Bacillus spp. on systemic immunity and intestinal health of weaned pigs infected with F18 E. coli. Weaned pigs (n = 36, 7.61 ± 0.40 kg BW) were randomly allotted to one of three treatments: a control diet and two diets supplemented with 500 mg/kg of Bacillus spp. strain 1 (PRO1) or strain 2 (PRO2). The experiment was conducted for 28 d, including 7 d before and 21 d after the first E. coli inoculation (d 0). The doses of F18 E. coli inoculum were 1010 cfu/3 mL oral dose daily for 3 consecutive days. Serum samples were collected on d 0 before inoculation, and d 3, 7, 14, and 21 post-inoculation (PI) to measure inflammatory meditators. All pigs were euthanized on d 21 PI to collect tissue samples for gut morphology, bacterial translocation, and gene expression analysis. Data were analyzed using the Mixed Procedure of SAS. Pigs in PRO1 had reduced (P < 0.05) serum haptoglobin on d 3 and d 6 PI compared with pigs in control. Supplementation of PRO1 or PRO2 reduced (P < 0.05) total coliforms in mesenteric lymph nodes on d 21 PI. Pigs in PRO1 had greater (P < 0.05) ileal villi height than pigs in control. Pigs in PRO2 had greater (P < 0.05) sulfomucin percentage in duodenal villi and greater (P < 0.05) sialomucin percentage in jejunal villi than pigs in control. Supplementation of PRO1 also up-regulated (P < 0.05) MUC2 gene expression in jejunal mucosa but reduced (P < 0.05) COX2 and IL1B gene expression in ileal mucosa on d 21 PI, compared with control. In conclusion, supplementation of Bacillus spp. may reduce systemic inflammation and enhance intestinal health of weaned pigs infected with F18 E. coli.


Metabolites ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 81
Author(s):  
Yijie He ◽  
Yanhong Liu ◽  
Peng Ji

This study explored the metabolomic profiles in ileal mucosa and colon digesta in response to enterotoxigenic Escherichia coli F18 (ETEC) infection and dietary use of probiotics and low-dose antibiotics. Weaned pigs (n = 48, 6.17 ± 0.36 kg body weight) were randomly allotted to one of four treatments. Pigs in the negative control (NC) were fed a basal diet without ETEC challenge, whereas pigs in the positive control (PC), antibiotic, and probiotic groups were fed the basal diet, basal diet supplemented with 50 mg/kg of carbadox, or 500 mg/kg of Bacillus subtilis, respectively, and orally challenged with ETEC F18. All pigs were euthanized at day 21 post-inoculation to collect ileal mucosa and colon digesta for untargeted metabolomic profiling using gas chromatography coupled with time-of-flight mass spectrometry. Multivariate analysis highlighted a more distinct metabolomic profile of ileal mucosa metabolites in NC compared to the ETEC-challenged groups. The relative abundance of 19 metabolites from the ileal mucosa including polyamine, nucleotide, monosaccharides, fatty acids, and organic acids was significantly different between the NC and PC groups (q < 0.1). In colon digesta, differential metabolites including 2-monoolein, lactic acid, and maltose were reduced in the carbadox group compared with the probiotics group. In conclusion, several differential metabolites and metabolic pathways were identified in ileal mucosa, which may suggest an ongoing intestinal mucosal repair in the ileum of ETEC-challenged pigs on day 21 post-inoculation.


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