scholarly journals Dietary Bacillus spp. enhanced growth and disease resistance of weaned pigs by modulating intestinal microbiota and systemic immunity

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Yijie He ◽  
Cynthia Jinno ◽  
Kwangwook Kim ◽  
Zhaohai Wu ◽  
Bie Tan ◽  
...  

Abstract Background Previous 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). Results Pigs in PRO1 (Bacillus subtilis DSM 32540) had greater (P < 0.05) body weight on d 7 and 14 PI, greater (P < 0.05) ADG from d 0 to 7 and d 7 to 14 PI, compared with pigs in CON (Control). Pigs in PRO1 had milder (P < 0.05) diarrhea on d 2 and 3 PI compared with pigs in CON. However, no differences were observed in growth performance and diarrhea score between PRO2 (Bacillus pumilus DSM 32539) and CON groups. Supplementation of PRO1 decreased (P < 0.05) lymphocyte counts on d 7 and 14 PI, compared with CON. Supplementation of PRO1 and PRO2 both reduced (P < 0.05) total coliforms in mesenteric lymph nodes on d 21 PI. Pigs in PRO2 had greater (P < 0.05) goblet cell number and sulfomucin percentage in duodenal villi and greater (P < 0.05) sialomucin percentage in jejunal villi than pigs in CON. Supplementation of PRO1 up-regulated (P < 0.05) MUC2 gene expression in jejunal mucosa and reduced (P < 0.05) PTGS-2 and IL1B gene expression in ileal mucosa on d 21 PI, compared with CON. Pigs in PRO1 had reduced (P < 0.05) relative abundance of families Lachnospiraceae, Peptostreptococcaceae and Pasteurellaceae in the ileum. Conclusions Supplementation of Bacillus subtilis DSM 32540 improved growth performance, alleviated diarrhea severity, enhanced gut health, and reduced systemic inflammation of weaned pigs infected with ETEC F18. Although Bacillus pumilus DSM 32539 was able to alleviate systemic inflammation, it had limited impacts on growth performance and severity of diarrhea of ETEC F18 challenged weaned pigs.

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.


2020 ◽  
Vol 98 (9) ◽  
Author(s):  
Yijie He ◽  
Kwangwook Kim ◽  
Lauren Kovanda ◽  
Cynthia Jinno ◽  
Minho Song ◽  
...  

Abstract 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 &lt; 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 &lt; 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 &lt; 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 &lt; 0.05) gene expression of CLDN1 in jejunal mucosa than pigs in the PC. Supplementation of carbadox or PRO reduced (P &lt; 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 &lt; 0.05) white blood cell number and neutrophil count, and serum haptoglobin concentration on day 7 PI, and less (P &lt; 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.


2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 76-77
Author(s):  
Kwangwook Kim ◽  
Yijie He ◽  
Xia Xiong ◽  
Amy Ehrlich ◽  
Xunde Li ◽  
...  

Abstract There is growing evidence to support the beneficial effects of supplementing direct-fed microbials (DFM) on performance, health status, and immune responses of weaned pigs. Therefore, the objective of this study was to investigate dietary supplementation of Bacillus subtilis (DSM 25841) on growth performance, diarrhea, gut permeability, immunity and metabolomic profiles of weaned pigs experimentally infected with a pathogenic F18 Escherichia coli (E. coli). E. coli infection reduced (P &lt; 0.05) growth performance and intestinal villi height, whereas increased (P &lt; 0.05) diarrhea and permeability in the jejunum compared with non-challenged control. Supplementation of Bacillus subtilis linearly enhanced average daily gain of E. coli infected pigs (d 0 to 5 post-inoculation (PI), P &lt; 0.05; d 0 to 11 PI, P = 0.058). Inclusion of high dose Bacillus subtilis reduced (P &lt; 0.05) jejunal permeability on d 5 and d 11 PI compared with the E. coli challenged control. E. coli challenged control pigs up-regulated (P &lt; 0.05) the mRNA expression of SLC5A10 and MUC2 on d 5 PI, but down-regulated (P &lt; 0.05) expression of SLC5A10, MUC2, and CLDN1 on d 11 PI in jejunal mucosa. Supplementation of Bacillus subtilis linearly up-regulated (P &lt; 0.05) the mRNA expression of CFTR and ZO1 on d 5 PI and SLC5A10 and MUC2 on d 11 PI in jejunal mucosa of E. coli infected pigs. E. coli infection increased (P &lt; 0.05) the mRNA expression of several immune genes in the ileal mucosa, while inclusion of Bacillus subtilis linearly down-regulated gene expression of IL1A on d 5 PI (P = 0.07) and IL6 on d 11 PI (P &lt; 0.05) in ileal mucosa of E. coli infected pigs. Supplementation of Bacillus subtilis modified (Fold change &gt; 1.5; FDR &lt; 0.20) metabolomic profiles in colon digesta, related to pathogenesis and amino acid metabolism. In conclusion, supplementation of Bacillus subtilis enhanced growth rate, improved gut health, and modified metabolomic profiles of weaned pigs experimentally infected with a pathogenic E. coli.


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.


2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 77-78
Author(s):  
Yi-Chi Cheng ◽  
Marcos E Duarte ◽  
Sung Woo Kim

Abstract The objective was to determine the functional and nutritional values of Corynebacterium glutamicum Cell Mass (CGCM) on growth performance and gut health of newly-weaned pigs. Forty newly-weaned pigs (21 d of age; initial BW 7.1 ± 0.4 kg) were allotted to 5 dietary treatments based on randomized complete block design with sex and BW as blocks. The lysine broth of CGCM (CJ Bio, Fort Dodge, IA) was homogenized by using French press and dried to obtain lysed CGCM. Dietary treatments were: basal diet with lysed CGCM at 0, 0.7, 1.4, 2.1%, and with 1.4% intact CGCM. Experimental diets were formulated based on nutrient requirements (NRC, 2012) and pigs were fed based on 2 phases (10 and 11 d for each phase). Titanium dioxide (0.4%) was added to phase 2 diets as an indigestible external marker to calculate nutrient digestibility. Feed intake and BW were measured at d 0, 10, and 21. Pigs were euthanized on d 21 to collect proximal and distal jejunal mucosa to measure TNF-α, IL-8, MDA, IgA, and IgG concentrations. Diets and ileal digesta were collected to measure AID. Data were analyzed by SAS using MIXED, REG, and GLM procedures. Overall, increasing daily lysed CGCM intake increased (P &lt; 0.05) ADG (211 to 296 g) and ADFI (432 to 501 g). Increasing levels of lysed CGCM decreased (P &lt; 0.05) MDA and changed (quadratic, P &lt; 0.05) IgA (max: 4.90 ng/mg at 1.13%) and IgG (max: 3.37 ng/mg at 1.04%) in the proximal jejunal mucosa. Increasing daily lysed CGCM intake had quadratic effect (P&lt; 0.05) of protein carbonyl (max: 6.3 μmol/mg at 4.9 g/d). Lysed CGCM potentially benefits growth performance and gut health of newly-weaned pigs by reducing oxidative stress and increasing immune response.


2020 ◽  
Vol 4 (3) ◽  
Author(s):  
Sangwoo Park ◽  
Jung Wook Lee ◽  
Kevin Jerez Bogota ◽  
David Francis ◽  
Jolie Caroline González-Vega ◽  
...  

Abstract This study was conducted to investigate the effects of a direct-fed microbial (DFM) product (Bacillus subtilis strain DSM 32540) in weaned pigs challenged with K88 strain of Escherichia coli on growth performance and indicators of gut health. A total of 21 weaned pigs [initial body weight (BW) = 8.19 kg] were housed individually in pens and fed three diets (seven replicates per diet) for 21 d in a completely randomized design. The three diets were a corn-soybean meal-based basal diet without feed additives, a basal diet with 0.25% antibiotics (neo-Oxy 10-10; neomycin + oxytetracycline), or a basal diet with 0.05% DFM. All pigs were orally challenged with a subclinical dose (6.7 × 108 CFU/mL) of K88 strain of E. coli on day 3 of the study (3 d after weaning). Feed intake and BW data were collected on days 0, 3, 7, 14, and 21. Fecal scores were recorded daily. On day 21, pigs were sacrificed to determine various indicators of gut health. Supplementation of the basal diet with antibiotics or DFM did not affect the overall (days 0–21) growth performance of pigs. However, antibiotics or DFM supplementation increased (P = 0.010) gain:feed (G:F) of pigs during the post-E. coli challenge period (days 3–21) by 23% and 24%, respectively. The G:F for the DFM-supplemented diet did not differ from that for the antibiotics-supplemented diet. The frequency of diarrhea for pigs fed a diet with antibiotics or DFM tended to be lower (P = 0.071) than that of pigs fed the basal diet. The jejunal villous height (VH) and the VH to crypt depth ratio (VH:CD) were increased (P &lt; 0.001) by 33% and 35%, respectively, due to the inclusion of antibiotics in the basal diet and by 43% and 41%, respectively due to the inclusion of DFM in the basal diet. The VH and VH:CD for the DFM-supplemented diet were greater (P &lt; 0.05) than those for the antibiotics-supplemented diet. Ileal VH was increased (P &lt; 0.05) by 46% due to the inclusion of DFM in the basal diet. The empty weight of small intestine, cecum, or colon relative to live BW was unaffected by dietary antibiotics or DFM supplementation. In conclusion, the addition of DFM to the basal diet improved the feed efficiency of E. coli-challenged weaned pigs to a value similar to that of the antibiotics-supplemented diet and increased jejunal VH and VH:CD ratio to values greater than those for the antibiotics-supplemented diet. Thus, under E. coli challenge, the test DFM product may replace the use of antibiotics as a growth promoter in diets for weaned pigs to improve feed efficiency and gut integrity.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 115-115
Author(s):  
M E Duarte ◽  
J Tyus ◽  
S W Kim

Abstract This study was to evaluate the supplemental effects of xylanase and probiotics on growth performance and gut health of newly weaned pigs. One hundred and forty-four pigs at 3 weeks of age with an initial body weight (BW) at 6.47 ± 0.03 kg were allotted (4 pigs per pen) to 3 dietary treatments based on a randomized complete block design with sex (gilts and barrow) and initial BW as blocks. The treatments were CON (no supplement), XYL (xylanase at 10,000 XU/kg, Xylamax, BRI, Durham, NC), and PRO (Bacillus spp. 6 x 109 CFU/kg plus xylanase 10,000 XU/kg, EnzaPro, BRI). All diets met the NRC (2012) requirements in 3 phases (P1 for 7 d, P2 for 14 d, and P3 for 14 d). The ADG, ADFI, and G/F were measured weekly. On d 21, 1 pig representing a median BW of each pen was euthanized to collect jejunal samples to measure the immune and oxidative stress status, microbiome, and histology. Data were analyzed using the MIXED procedure of SAS. Treatment was a fixed effect and blocks were random effects. There were no fixed effects among treatments on growth performance, immune and oxidative stress status, and histology. Use of xylanase increased (P &lt; 0.05) the Chao1 index in jejunal mucosa of pigs (24.6 to 29.5). Relative abundance of the Enterobacteriaceae in jejunal mucosa tended to decrease (P = 0.083) with the use of xylanase (16.2 to 1.6%). The PRO increased (P &lt; 0.05) the relative abundance of Mitsuokella compared with others treatments (0.9 vs. 4.8%). In conclusion, supplementation of xylanase or Mitsuokella can enhance the microbiome in jejunal mucosa by increasing the diversity and beneficial bacterial abundance, whereas by reducing pathogenic bacterial abundance without affecting the growth performance, histology, immune, and oxidative stress status.


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.


2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 133-134
Author(s):  
M E Duarte ◽  
E Grilli ◽  
S W Kim

Abstract This study was to determine the supplemental effects of lauric acid and a microencapsulated blend of organic acids and botanicals (AviPlusS, Vetagro, Chicago, IL) on growth performance and gut health in newly weaned pigs. Ninety-six pigs (48 barrows and 48 gilts, 21 d-of-age, 6.5 ± 0.4 kg BW) were randomly allotted (3 pigs/ pen) to 4 treatments in RCBD arranged by 2 factors: lauric acid (0 or 0.1%) and AviPlusS (0 or 0.3%). Initial BW and sex were blocks. Diets met the NRC (2012) requirements, medicated, included ZnO, and prepared in 3 phases (phase 1 for 7 d; phase 2 for 11 d; and phase 3 for 14 d). At the end of each phase, ADG, ADFI, and G:F was measured. On d 32, 1 pig representing the median BW of each pen was euthanized to collect jejunal mucosa to analyze immune and oxidative stress status and microbiome. Data were analyzed using the MIXED procedure of SAS. Two factors were fixed effects and blocks were random effects. Overall performance was not affected by 2 factors. Combinational use of lauric acid and AviPlusS tended to increase (P = 0.053) the relative abundance of Lactobacillaceae (18.11 vs. 9.59 and 10.26%). Lauric acid tended to reduce (P = 0.081) the relative abundance of Methylobacteriaceae (10.34 to 6.30%). Lauric acid tended to increase (P = 0.083) the relative abundance of Veillonellaceae (2.05 to 9.28%) which tended to be reduced (P = 0.078) when combined with AviPlusS. Combinational use of lauric acid and AviPlusS increased (P &lt; 0.05) the relative abundance of Lactobacillus (23.34 vs. 13.21 and 17.56%) compared with the use of lauric acid or AviPlusS alone. Collectively, lauric acid can modulate the microbiome to enhance gut health, which can be further enhanced when combined with AviPlusS.


2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 77-77
Author(s):  
Yi-Chi Cheng ◽  
Marcos E Duarte ◽  
Sung Woo Kim

Abstract The objective was to test the efficacy of L-Thr or L-Val with biomass (ThrPro or ValPro; CJ BIO, Fort Dodge, IA) on growth performance and health compared with L-Thr or L-Val (purified) in newly-weaned pigs. There were 9 dietary treatments based on NRC (2012) requirements: NC (basal diet meeting 70% of requirements for SID Thr and Val); 4 treatments based on basal diet meeting 95% of requirements: LT (with L-Thr), LV (with L-Val), PT (with ThrPro), and PV (with ValPro); HLT (fivefold L-Thr in LT), HLV (fivefold L-Val in LV), HPT (fivefold ThrPro in PT), and HPV (fivefold ValPro in PV). Pigs were fed for 26 d in 2 phases. Feed intake and BW were measured at d 0, 5, 10, 17, and 26. Blood was collected on d 21 for liver function analysis. Pigs were euthanized on d 26 to collect jejunal mucosa for gut health parameters. Data were analyzed by SAS using MIXED procedure. Pigs fed LT and LV or PT and PV had higher (P &lt; 0.05) ADG than pigs fed NC during phase 2. Pigs fed LT, LV, PT, and PV had higher ADFI and G:F than pigs fed NC during d 17 to 26. Pigs fed PT and PV had lower (P &lt; 0.05) G:F than pigs fed HPT and HPV during phase 2. Pigs fed LT and LV or PT and PV had lower (P &lt; 0.05) BUN than pigs fed NC. Overall, pigs fed PT and PV had no differences in any analyzed measurements from pigs fed LT and LV and pigs fed HPT and HPV. In conclusion, PT and PV were equally efficient to LT and LV for growth performance and health. HPT and HPV did not impair liver and gut health compared with PT and PV.


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