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 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.

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 &lt; 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 &lt; 0.05) abundant but Firmicutes were less (P &lt; 0.05) abundant in ileal digesta of pigs fed with antibiotics than pigs in probiotics. Firmicutes were more (P &lt; 0.05) abundant in colon and feces of NC than of antibiotics. Bifidobacterium was least (P &lt; 0.05) abundant throughout all sites and Prevotella 1 was most (P &lt; 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 &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.

scholarly journals Ameliorative Effect of Bacillus subtilis on Growth Performance and Intestinal Architecture in Broiler Infected with Salmonella

Animals ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 190 ◽  
Author(s):  
Alaeldein M. Abudabos ◽  
Muttahar H. Ali ◽  
Mohammed A. Nassan ◽  
Ahmad A. Saleh
Keyword(s):  
Bacillus Subtilis ◽  
Basal Diet ◽  
Positive Control ◽  
Negative Control ◽  
Broiler Chicks ◽  
Feed Conversion ◽  
Negative Effects ◽  
Villus Height ◽  
Bacillus Subtilis Atcc ◽  
Ameliorative Effect

A total of 600 day-old broiler chicks (Ross 308) confirmed for the absence of Salmonella were randomly allocated to five treatments each with 10 replicates: negative control (basal diet only); positive control (basal diet) + infected with Salmonella; T1, Salmonella infected + avilamycin; T2, Salmonella infected + Bacillus subtilis (ATCC PTA-6737; 2 × 107 CFU/g) and T3, Salmonella infected + B. subtilis (DSM 172999; 1.2 × 106 CFU/g). The results revealed that feed intake (FI) and body weight (BW) were significantly (p < 0.01) lower in T1 compared to T2. The feed conversion ratio (FCR) was significantly (p < 0.01) lower in T2 and T3 compared to other treatments. Similarly, the performance efficiency factor (PEF) was also significantly (p < 0.01) higher in T2 and T3 compared to positive control. Villus height was significantly (p < 0.01) higher in T2 compared to all other treatments. However, villus width and surface area were significantly (p < 0.01) higher in T1. In conclusion, dietary supplementation with B. subtilis improved growth and intestinal health by reversing the negative effects of Salmonellosis.

scholarly journals Efficacy of organic acids as an alternative to antibiotic growth promoters in weaned pigs

2018 ◽  
Vol 17 (06) ◽  
pp. 8-14
Author(s):  
Tung M. Che
Keyword(s):  
Organic Acids ◽  
Antibiotic Treatment ◽  
Average Daily Gain ◽  
Basal Diet ◽  
Positive Control ◽  
Negative Control ◽  
Growth Promoters ◽  
Weaned Pigs ◽  
E Coli ◽  
Diarrhea Incidence

The objective of the experiment was to compare effects of dietary supplementation of organic acids (ProHacid Advance, PRO) and antibiotics on growth performance, diarrhea incidence, frequency of antibiotic treatment, and E. coli shedding in weaned pigs. A total of 224 crossbred weaned pigs [(Yorkshire x Landrace) x Duroc; 29 days old)] were allotted to 1 of 4 treatments with 7 replicate pens of 8 pigs each according to sex, litter origin and weight in an RCBD. The 4 dietary treatments included (1) basal diet + 50 mg/kg neomycin and 10 mg/kg enramycin (positive control, PC), (2) basal diet without antibiotics (negative control, NC), (3) As 2 + 0.25% PRO, and (4) As 2 + 0.5% PRO. Over a 4-week study, the results showed that there were no differences in the average daily gain and average daily feed intake of pigs among the 4 treatments (P = 0.328). However, pigs fed the PC diet (1.642) and 0.25% PRO-supplemented diet (1.641) had a lower feed to gain ratio (P < 0.05) than those fed the NC diet (1.808). The diarrhea incidence of pigs fed the 0.25% PRO-supplemented diet (7.36%) was lower (P < 0.01) than that of pigs fed the PC diet (11.61%), NC diet (16.70%), and 0.5% PRO-supplemented diet (10.08%). The frequency of antibiotic treatment of pigs consuming the 0.25% PRO-supplemented diet (4.67%) was lower (P < 0.01) than that of pigs consuming the PC diet (7.33%), NC diet (9.53%) and 0.5% PRO-supplemented diet (7.65%). No differences were found in the number of fecal E. coli among the 4 treatments. In brief, 0.25% PRO added to a nursery pig diet would be considered a potential alternative to the use of antibiotics in feed, but further research on this aspect is needed

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

2021 ◽  
Author(s):  
Jinyoung Lee ◽  
Jong Woong Kim ◽  
Heidi N. Hall ◽  
Charles M. Nyachoti
Keyword(s):  
Growth Performance ◽  
Phase 1 ◽  
Basal Diet ◽  
Positive Control ◽  
Negative Control ◽  
Nutrient Digestibility ◽  
Feed Ratio ◽  
Weaned Pigs ◽  
Gut Morphology ◽  
Two Phases

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

scholarly journals PSVIII-2 Dietary supplementation of Bacillus subtilis altered metabolites in the intestine of weaned pigs

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 208-208
Author(s):  
Yijie He ◽  
Kwangwook Kim ◽  
Cynthia Jinno ◽  
Xunde Li ◽  
Yanhong Liu
Keyword(s):  
Bacillus Subtilis ◽  
Statistical Significance ◽  
Control Diet ◽  
Dietary Supplementation ◽  
Enterotoxigenic Escherichia Coli ◽  
P Value ◽  
Weaned Pigs ◽  
Ileal Mucosa ◽  
Galactose Metabolism ◽  
E Coli

Abstract This experiment aimed to investigate the effects of dietary supplementation of Bacillus subtilis probiotics on metabolites in the intestines of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (E. coli). Forty-eight weaned pigs (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 days with 7 days before and 21 days after the first E. coli inoculation. The F18 E. coli were given to pigs at 1010 CFU/3 mL dose for three consecutive days. At the end of the experiment, all pigs were euthanized to collect ileal mucosa and colon digesta for the analysis of metabolomic profiles by gas chromatography time of flight-mass spectrometer (GCTOF-MS). All data were analyzed by an online MetaboAnalyst tool (https://www.metaboanalyst.ca/). Statistical significance was declared at P &lt; 0.05 and the false discovery rate–adjusted P value (q value) &lt; 0.20. A total of 282 (141 identified and 121 unidentified) and 196 (127 identified and 69 unidentified) metabolites were detected in ileal mucosa and colon digesta, respectively. Forty-nine identified metabolites in ileal mucosa significantly differed among experimental groups (P &lt; 0.05; q &lt; 0.20). The most impacted metabolic pathways were galactose metabolism, aspartate and glutamate metabolism, fructose and mannose degradation, pentose phosphate pathway, and urea cycle. However, in colon digesta, only 7 identified metabolites differed among experimental groups (P &lt; 0.05; q &lt; 0.20) and the majority of them were involved in purine metabolism. Results of metabolomics indicated that supplementation of Bacillus subtilis or antibiotics altered metabolites in the intestines of weaned pigs. In particular, more treatment impacts were observed in the metabolite profiles in ileal mucosa compared with colon digesta.

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 &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.

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.

scholarly journals Growth Performance, Serum Biochemical Indices, Duodenal Histomorphology, and Cecal Microbiota of Broiler Chickens Fed on Diets Supplemented with Cinnamon Bark Powder at Prestarter and Starter Phases

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 94
Author(s):  
Mohammed M. Qaid ◽  
Saud I. Al-Mufarrej ◽  
Mahmoud M. Azzam ◽  
Maged A. Al-Garadi ◽  
Hani H. Albaadani ◽  
...  
Keyword(s):  
Body Weight ◽  
Broiler Chickens ◽  
Body Weight Gain ◽  
Basal Diet ◽  
Positive Control ◽  
Negative Control ◽  
Broiler Chicks ◽  
Feed Conversion ◽  
Serum Globulin ◽  
Maximum Improvement

Ross 308 broiler chicks (n = 240) aged 1 day were assigned to five groups for eight replicates (six chicks for each) (3♂ and 3♀). Basal dietary groups were supplemented by 2000, 4000, and 6000 mg/kg cinnamon (CN) for 21 days. Basal diet alone was used as a negative control, and basal antibiotic diet (Colimox) was used as a positive control. At 10, 14, and 21 days of age, chicks that received 2000 mg CN and Colimox had a higher body weight, resulting in an increase in body weight gain. CN also resulted in the maximum improvement in the feed conversion ratio and feed efficiency over 1–21 days at the level of 2000 mg/kg. At days 10, the maximum relative breast weight was 2000 mg/kg of CN. Mean serum albumin concentrations, duodenal villus height, and goblet cell density increased (p < 0.05) by 2000 mg/kg of CN, and mean serum globulin and total protein concentrations and crypt depth increased (p < 0.05) by 6000 mg/kg of CN compared with control. Increased cecal Escherichia coli number was CN dose-dependent. In conclusion, dietary inclusion of 2000 mg/kg CN can be applied as an alternative to in-feed antibiotics for broiler starter diet.

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