scholarly journals PSIX-5 Dietary supplementation of Bacillus subtilis modified fecal microbiome of weaned pigs in comparison to antibiotics

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 183-183
Author(s):  
Cynthia Jinno ◽  
Yijie He ◽  
Xunde Li ◽  
Yanhong Liu
Keyword(s):  
Bacillus Subtilis ◽  
Relative Abundance ◽  
Hypervariable Region ◽  
Negative Control ◽  
Rrna Gene ◽  
Post Inoculation ◽  
Weaned Pigs ◽  
Fecal Samples ◽  
Fecal Microbiome ◽  
E Coli

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 < 0.05) in feces collected on d 0 before E. coli inoculation and lowest (P < 0.05) on d -7 feces. Pigs supplemented with DFM had lower (P < 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 < 0.05) abundant on d -7 and lowest (P < 0.05) on d 21 PI. However, Actinobacteria and Firmicutes were most (P < 0.05) abundant on d 21 PI. Pigs in the NC and DFM groups had greater (P < 0.05) relative abundance of Firmicutes than pigs fed with antibiotics on d 0 and 7. Supplementation of antibiotics reduced (P < 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.

PSII-15 Dietary Supplementation of Bacillus Subtilis Modified Intestinal Microbiome of Weaned Pigs Differently to Antibiotics

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 161-162
Author(s):  
Cynthia Jinno ◽  
Yijie He ◽  
Yanhong Liu
Keyword(s):  
Bacillus Subtilis ◽  
Hypervariable Region ◽  
Alpha Diversity ◽  
Basal Diet ◽  
Positive Control ◽  
Negative Control ◽  
Intestinal Microbiome ◽  
Weaned Pigs ◽  
E Coli ◽  
Downstream Analysis

Abstract 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.

scholarly journals PSVIII-7 Effects of Bacillus subtilis on colon digesta metabolomic profiles in weanling pigs experimentally infected with a pathogenic E. coli

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 203-203
Author(s):  
Kwangwook Kim ◽  
Yanhong Liu
Keyword(s):  
Bacillus Subtilis ◽  
Positive Control ◽  
Negative Control ◽  
High Dose ◽  
Post Inoculation ◽  
Weaned Pigs ◽  
E Coli ◽  
Gut Barrier Function ◽  
Pathogenic Escherichia Coli ◽  
Lysine Degradation

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 > 1.5; FDR < 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.

scholarly journals Fecal microbiome profiles of neonatal dairy calves with varying severities of gastrointestinal disease

PLoS ONE ◽  
2022 ◽  
Vol 17 (1) ◽  
pp. e0262317
Author(s):  
Giovana S. Slanzon ◽  
Benjamin J. Ridenhour ◽  
Dale A. Moore ◽  
William M. Sischo ◽  
Lindsay M. Parrish ◽  
...  
Keyword(s):  
Health Status ◽  
Relative Abundance ◽  
Gastrointestinal Disease ◽  
Bifidobacterium Longum ◽  
Dairy Calves ◽  
Rrna Gene ◽  
Probiotic Properties ◽  
Fecal Samples ◽  
Fecal Microbiome ◽  
Microbiome Composition

Gastrointestinal disease (GI) is the most common illness in pre-weaned dairy calves. Studies have associated the fecal microbiome composition with health status, but it remains unclear how the microbiome changes across different levels of GI disease and breeds. Our objective was to associate the clinical symptoms of GI disease with the fecal microbiome. Fecal samples were collected from calves (n = 167) of different breeds (Holstein, Jersey, Jersey-cross and beef-cross) from 4–21 d of age. Daily clinical evaluations assessed health status. Calves with loose or watery feces were diagnosed with diarrhea and classified as bright-sick (BS) or depressed-sick (DS) according to behavior. Calves with normal or semiformed feces and no clinical illness were classified as healthy (H). One hundred and three fecal samples were obtained from consistently healthy calves and 64 samples were from calves with diarrhea (n = 39 BS; n = 25 DS). The V3-V4 region of 16S rRNA gene was sequenced and analyzed. Differences were identified by a linear-mixed effects model with a negative binomial error. DS and Jersey calves had a higher relative abundance of Streptococcus gallolyticus relative to H Holstein calves. In addition, DS calves had a lower relative abundance of Bifidobacterium longum and an enrichment of Escherichia coli. Species of the genus Lactobacillus, such as an unclassified Lactobacillus, Lactobacillus reuteri, and Lactobacillus salivarius were enriched in calves with GI disease. Moreover, we created a model to predict GI disease based on the fecal microbiome composition. The presence of Eggerthella lenta, Bifidobacterium longum, and Collinsella aerofaciens were associated with a healthy clinical outcome. Although lactobacilli are often associated with beneficial probiotic properties, the presence of E. coli and Lactobacillus species had the highest coefficients positively associated with GI disease prediction. Our results indicate that there are differences in the fecal microbiome of calves associated with GI disease severity and breed specificities.

scholarly journals 140 Effects of Bacillus subtilis probiotics on growth performance, diarrhea, and fecal β-hemolytic coliforms of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 79-80
Author(s):  
Yijie He ◽  
Kwangwook Kim ◽  
Cynthia Jinno ◽  
Lauren Kovanda ◽  
Seijoo Yang ◽  
...  
Keyword(s):  
Body Weight ◽  
Bacillus Subtilis ◽  
Growth Performance ◽  
Feed Efficiency ◽  
Control Diet ◽  
Positive Control ◽  
Negative Control ◽  
Enterotoxigenic Escherichia Coli ◽  
Weaned Pigs ◽  
E Coli

Abstract The objective of this experiment was to investigate the effects of Bacillus subtilis on growth performance, diarrhea and fecal β-hemolytic coliforms of weaned pigs experimentally infected with a strain of E. coli (F18, express genes of LT, STb, and SLT 2 toxins). Weaned pigs (n = 48, 6.17 ± 0.36 kg 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 or 500 mg/kg of Bacillus subtilis probiotics. The experiment lasted 28 d with 7 d before and 21 d after the first E. coli inoculation. The F18 E. coli were given to pigs at 1010 CFU/3 mL dose for three consecutive d. Diarrhea score was daily recorded for each pig to calculate frequency of diarrhea. Fecal samples were collected on d 0, 3, 7, 14, and 21 PI to analyze β-hemolytic coliforms. Data were analyzed using the Mixed Procedure of SAS. Pigs supplemented with carbadox had greater (P < 0.05) body weight on d 7, 14, and 21 PI than pigs in the PC and probiotics group. Supplementation of probiotics enhanced pig body weight on d 21 PI, compared with the PC. E. coli challenge reduced (P < 0.05) ADG and feed efficiency from d 0 to 21 PI, while supplementation of antibiotics or probiotics enhanced ADG and feed efficiency from d 0 to 21 PI. Pigs in carbadox and probiotics groups had reduced (P < 0.05) frequency of diarrhea throughout the experiment and fecal β-hemolytic coliforms on d 7 PI than pigs in the PC. In conclusion, supplementation of Bacillus subtilis could enhance disease resistance and promote growth performance of weaned pigs under disease challenge condition.

scholarly journals Metabolomic Profile of Weaned Pigs Challenged with E. coli and Supplemented with Carbadox or Bacillus subtilis

Metabolites ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 81
Author(s):  
Yijie He ◽  
Yanhong Liu ◽  
Peng Ji
Keyword(s):  
Bacillus Subtilis ◽  
Basal Diet ◽  
Positive Control ◽  
Negative Control ◽  
Enterotoxigenic Escherichia Coli ◽  
Post Inoculation ◽  
Metabolomic Profile ◽  
Weaned Pigs ◽  
Ileal Mucosa ◽  
Flight Mass Spectrometry

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.

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

2021 ◽  
Author(s):  
Kwangwook Kim ◽  
Cynthia Jinno ◽  
Peng Ji ◽  
Yanhong Liu
Keyword(s):  
Relative Abundance ◽  
Metabolic Pathways ◽  
Recommended Dose ◽  
Control Group ◽  
Metabolic Profiles ◽  
Post Inoculation ◽  
Metabolomic Profile ◽  
Weaned Pigs ◽  
E Coli ◽  
Etec Infection

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.

scholarly journals 141 Supplementation of Bacillus subtilis modified fecal microbiota of weaning pigs experimentally infected with pathogenic E. coli

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 81-81
Author(s):  
Cynthia Jinno ◽  
Kwangwook Kim ◽  
Minho Song ◽  
Peng Ji ◽  
Elizabeth Maga ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Compositional Data ◽  
Diversity Index ◽  
Fecal Microbiota ◽  
Rrna Gene ◽  
Post Inoculation ◽  
Double Dose ◽  
E Coli ◽  
Shannon Diversity ◽  
D 12

Abstract The objective of this experiment was to investigate the influence of Bacillus subtilis supplements on fecal microbiota of pigs experimentally infected with F-18 E. coli. Forty-eight pigs (6.73 ± 0.77 kg BW) were individually housed in disease containment rooms and randomly allotted to one of four treatments with 12 replicate pigs per treatment. Four treatments included negative control (NC), positive control (PC), single dose probiotics, and double dose probiotics. Pigs in the NC and PC groups were fed with basal diet but without or with E. coli challenge, respectively. Pigs in the probiotics groups were fed the diets either supplemented with 1.28 × 109 CFU or 2.56 × 109 CFU Bacillus subtilis/kg feed and challenged with E. coli. The experiment lasted 18 d with 7 d before and 11 d after the first E. coli inoculation (d 0). Fecal samples were collected on d 5 and d 12 post inoculation (PI) and fecal microbiota was analyzed using 16S rRNA gene sequencing at the V4 hypervariable region and compositional data was analyzed using QIIME2 (2016.6). Pigs in the NC group had less (P < 0.05) Shannon diversity index than pigs in the other groups on d 5 PI, but no difference was observed in Shannon diversity among all treatment groups on d 12 PI. Bray Curtis PCoA analysis displayed that pigs in both probiotics group were separately clustered from the NC and PC groups on d 5 PI. Supplementation of single or double dose probiotics reduced (P < 0.05) the abundance of fecal Firmicutes compared with NC and PC. Within this phylum, the abundance of Lachnospiraceae was increased (P < 0.05), but the abundance of Lactobacillaceae and Streptococcaceae was decreased (P < 0.05). In conclusion, supplementation of Bacillus subtilis could modify gut microbiota of weaned pigs challenged with F18 E. coli.

scholarly journals PSIV-11 Effects of very low-dose antibiotics on gene expression profiles in ileal mucosa of weaned pigs infected with a pathogenic E. coli

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 286-286
Author(s):  
Kwangwook Kim ◽  
Sungbong Jang ◽  
Yanhong Liu
Keyword(s):  
Gene Expression ◽  
Systemic Inflammation ◽  
Low Dose ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Post Inoculation ◽  
Growth Promoter ◽  
Weaned Pigs ◽  
Ileal Mucosa ◽  
E Coli

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.

187 Effects of Oligosaccharide-based Polymer on Growth Performance, Diarrhea, and Fecal β-hemolytic Coliforms in Weanling Pigs Experimentally Infected with a Pathogenic E. coli

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 90-90
Author(s):  
Kwangwook Kim ◽  
Yijie He ◽  
Cynthia Jinno ◽  
Seijoo Yang ◽  
Xunde Li ◽  
...  
Keyword(s):  
Growth Performance ◽  
Feed Efficiency ◽  
Basal Diet ◽  
Watery Diarrhea ◽  
Experimental Unit ◽  
Post Inoculation ◽  
Weaned Pigs ◽  
E Coli ◽  
Diet Control ◽  
Dietary Treatments

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.

PSV-B-18 Characterization of the fecal microbiome of healthy adult beagle dogs supplemented with graded levels of bacillus coagulans GBI-30, 6086

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 337-338
Author(s):  
Heather L Acuff ◽  
Tara N Gaire ◽  
Tyler Doerksen ◽  
Andrea Lu ◽  
Michael P Hays ◽  
...  
Keyword(s):  
Relative Abundance ◽  
Fixed Effects ◽  
Mixed Model ◽  
Healthy Adult ◽  
Random Effect ◽  
Bacillus Coagulans ◽  
Community Diversity ◽  
Rrna Gene ◽  
Beagle Dogs ◽  
Fecal Microbiome

Abstract This study aimed to evaluate the effect of Bacillus coagulans GBI-30, 6086 on the fecal microbiome of healthy adult dogs. Extruded diets containing graded levels of probiotic applied either to the base ration before extrusion or as a topical coating post-extrusion were randomly assigned to ten individually-housed Beagle dogs (7 castrated males, 3 spayed females) of similar age (5.75 ± 0.23 yr) and body weight (12.3 ± 1.5 kg) in a 5 x 5 replicated Latin square with 16-d adaptation and 5-d total fecal collection for each period. Five dietary treatments were formulated to deliver a dose of 0-, 6-, 7-, 8-, or 9-log10 CFU·dog-1·d-1. Fresh fecal samples (n=50) were analyzed by 16S rRNA gene pyrosequencing. Community diversity was evaluated in R (v4.0.3, R Core Team, 2019). Relative abundance data were analyzed using a mixed model (v9.4, SAS Institute, Inc., Cary, NC) with treatment and period as fixed effects and dog as a random effect. Results were considered significant at P &lt; 0.05. Predominant phyla were Firmicutes (mean 81.2% ± 5), Actinobacteria (mean 9.9% ± 4.4), Bacteroidetes (mean 4.5% ± 1.7), Proteobacteria (mean 1.3% ± 0.7), and Fusobacteria (mean 1.1% ± 0.6). No evidence of shifts in predominant phyla, class, family, or genus taxonomic levels were observed except for the Bacillus genus, which had a greater relative abundance (P = 0.0189) in the low probiotic coating and high probiotic coating treatment groups compared to the extruded probiotic group. Alpha-diversity indices (Richness, Chao1, ACE, Shannon, Simpson, Inverse Simpson, and Fisher) and beta-diversity metrics (principal coordinate analysis and multi-dimensional scaling) were similar for all treatments. This data indicates that supplementation with Bacillus coagulans GBI-30, 6086 at a dose of up to 9 log10 CFU·d-1 did not alter the overall diversity of the fecal microbiome of healthy adult dogs over a 21-d period.

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