Dietary Administration Effects of Exopolysaccharide Produced by Bacillus tequilensis PS21 Using Riceberry Broken Rice, and Soybean Meal on Growth Performance, Immunity, and Resistance to Streptococcus agalactiae of Nile tilapia (Oreochromis niloticus)
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Bacterial Cultivation
2.2. Bacterial EPS Production
2.3. Crude EPS Extraction and Compositional Analysis
2.4. Monosaccharide Composition Analysis
2.5. Fourier-Transform Infrared Spectroscopy (FTIR) Analysis
2.6. Scanning Electron Microscopic (SEM) Analysis
2.7. Antioxidant Activity Assays
2.7.1. Scavenging Ability on 2,2-Diphenyl-1-picrylhydrazyl (DPPH•) Free Radicals
2.7.2. Ferric Ion Antioxidant Reducing Power (FRAP)
2.7.3. Scavenging Ability on Hydroxyl Radicals
2.8. Antimicrobial Activity of EPS
2.9. Trial Fish Diet Formula
2.10. Experimental Design
2.11. Sample Collections
2.12. Growth Measurements
2.13. Body Chemical Composition and Organosomatic Indices
2.14. Blood Chemical Analysis and Antioxidant Enzyme Activity
2.15. Hematological Parameters
2.16. Immunological Analysis
2.17. Serum Bactericidal Activity
2.18. Gene Expression
2.19. Challenge Test
2.20. Statistical Analysis
3. Results
3.1. EPS Characterization
3.2. EPS Bioactivities
3.3. Growth Performances
3.4. Hematological and Blood Chemical Profiles
3.5. Body Composition and Organosomatic Indices
3.6. Blood Biochemical Profiles and Immunological/Antioxidant Parameters
3.7. Gene Expressions in the Spleen and Liver
3.8. Bactericidal Activity
3.9. Pathogen Challenge Test
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Accession No. | Primer Sequence (5′→3′) | Tm (°C) | Product Size (bp) |
---|---|---|---|---|
β-actin | EF206801 | Forward: GCTACTCCTTCACCACCACAG Reverse: CGTCAGGCAGCTCGTAACTC | 58 | 138 |
IL-1β | JF957370 | Forward: TGCACTGTCACTGACAGCCAA Reverse: ATGTTCAGGTGCACTTTGCGG | 58 | 144 |
TNF-α | AY428948 | Forward: CCAGAAGCACTAAAGGCGAAGA Reverse: CCTTGGCTTTGCTGCTGATC | 58 | 82 |
Sample | Content (mg/g EPS) | EPS Production (g/100 mL) | ||
---|---|---|---|---|
Carbohydrate | Protein | Nucleic Acid | ||
EPS | 404.23 ± 9.86 a (76.58%) | 15.66 ± 0.61 c (2.97%) | 107.92 ± 4.58 b (20.45%) | 2.40 ± 0.20 |
Sample | DPPH Radical Scavenging Activity (%) | FRAP Value (mg FeSO4/g DW) | Hydroxyl Radical Scavenging Activity (%) | Inhibition Zone (mm Diameter) |
---|---|---|---|---|
EPS (20 mg/mL) | 65.50 ± 0.31 b | 2.07 ± 0.04 a | 80.53 ± 0.87 a | 14.17 ± 0.76 b |
Dextran (20 mg/mL) | 73.98 ± 0.27 a | 2.23 ± 0.04 a | 80.94 ± 0.46 a | 15.50 ± 0.05 a |
Chemical Composition | T1 (0.0 g/kg) | T2 (0.1 g/kg) | T3 (0.2 g/kg) | T4 (1.0 g /kg) | T5 (2.0 g/kg) |
---|---|---|---|---|---|
Crude protein (%) | 32.68 | 32.16 | 32.38 | 31.97 | 32.75 |
Crude lipid (%) | 4.13 | 4.17 | 4.29 | 4.74 | 4.31 |
Crude fiber (%) | 4.47 | 4.35 | 4.29 | 4.61 | 4.48 |
Moisture (%) | 12.23 | 12.95 | 12.82 | 13.08 | 12.73 |
Ash (%) | 10.09 | 9.80 | 9.87 | 10.17 | 9.98 |
NFE (%) 1 | 48.63 | 49.53 | 49.18 | 48.51 | 48.47 |
GE (MJ/kg) 2 | 17.71 | 17.75 | 17.79 | 17.76 | 17.77 |
Parameter | T1 (0.0 g/kg) | T2 (0.1 g/kg) | T3 (0.2 g/kg) | T4 (1.0 g/kg) | T5 (2.0 g/kg) |
---|---|---|---|---|---|
Initial body weight (IBW; g) | 15.23 ± 0.73 | 14.90 ± 0.34 | 15.20 ± 0.95 | 15.16 ± 1.19 | 15.20 ± 1.01 |
Initial length (IL; cm) | 7.63 ± 0.15 | 7.56 ± 0.23 | 7.66 ± 0.11 | 7.73 ± 0.30 | 7.60 ± 0.17 |
Final body weight (FBW; g) | 57.61 ± 2.73 | 57.81 ± 3.29 | 60.01 ± 3.03 | 61.75 ± 1.37 | 60.22 ± 2.20 |
Final length (FL; cm) | 11.40 ± 0.09 | 11.41 ± 0.26 | 11.60 ± 0.43 | 11.73 ± 0.12 | 11.76 ± 0.20 |
Weight gain (WG; g) | 42.39 ± 2.19 | 42.90 ± 3.08 | 44.83 ± 2.30 | 46.60 ± 1.67 | 45.00 ± 1.33 |
Length gain (LG; cm) | 3.77 ± 0.15 | 3.83 ± 0.05 | 3.90 ± 0.34 | 4.00 ± 0.36 | 4.20 ± 0.10 |
Average daily growth gain (ADG; g/day) | 0.68 ± 0.03 | 0.69 ± 0.05 | 0.72 ± 0.04 | 0.75 ± 0.03 | 0.73 ± 0.02 |
Specific growth rate; SGR (%/day) | 2.14 ± 0.05 | 2.18 ± 0.07 | 2.22 ± 0.05 | 2.26 ± 0.13 | 2.21 ± 0.06 |
Feed conversion ratio; (FCR) | 1.46 ± 0.03 | 1.51 ± 0.06 | 1.53 ± 0.04 | 1.47 ± 0.11 | 1.52 ± 0.20 |
Survival rate (%) | 83.33 ± 6.66 | 88.88 ± 6.94 | 85.55 ± 6.93 | 92.22 ± 6.94 | 88.89 ± 10.71 |
Parameter | T1 (0.0 g/kg) | T2 (0.1 g/kg) | T3 (0.2 g/kg) | T4 (1.0 g/kg) | T5 (2.0 g/kg) |
---|---|---|---|---|---|
RBC (×106 cells/mm3) | 2.10 ± 0.06 | 1.96 ± 0.11 | 2.10 ± 0.16 | 2.11 ± 0.29 | 2.02 ± 0.13 |
Hemoglobin (g/dL) | 11.00 ± 2.98 | 9.53 ± 3.30 | 10.50 ± 1.73 | 11.66 ± 1.62 | 10.73 ± 0.66 |
Haematocrit (%) | 33.00 ± 9.64 | 27.66 ± 9.50 | 31.00 ± 6.24 | 34.66 ± 4.16 | 31.33 ± 1.52 |
WBC (×103 cells/mm3) | 5.82 ± 1.46 | 4.14 ± 0.82 | 5.91 ± 1.80 | 6.39 ± 3.31 | 6.55 ± 1.10 |
Neutrophils (%) | 30.00 ± 5.29 b | 34.00 ± 1.58 b | 32.66 ± 7.50 ab | 38.66 ± 1.52 a | 39.00 ± 1.73 a |
Eosinophils (%) | 1.33 ± 0.57 | 1.16 ± 0.28 | 1.00 ± 0.01 | 1.16 ± 0.76 | 1.33 ± 0.57 |
Lymphocytes (%) | 62.00 ± 4.58 | 63.66 ± 7.09 | 65.00 ± 5.19 | 66.66 ± 5.85 | 68.66 ± 2.51 |
Monocytes (%) | 2.66 ± 0.57 | 3.66 ± 1.52 | 4.66 ± 1.52 | 3.00 ± 1.00 | 4.00 ± 0.01 |
MCV (fL) | 175.00 ± 1.00 | 169.00 ± 4.35 | 171.33 ± 1.52 | 168.66 ± 6.80 | 174.66 ± 2.51 |
MCH (pg) | 54.96 ± 0.80 | 57.33 ± 5.00 | 57.76 ± 2.08 | 57.00 ± 1.01 | 57.50 ± 2.52 |
MCHC (g/dL) | 31.43 ± 0.65 | 33.86 ± 2.11 | 33.70 ± 1.15 | 33.86 ± 0.90 | 32.86 ± 1.35 |
Parameter | T1 (0.0 g/kg) | T2 (0.1 g/kg) | T3 (0.2 g/kg) | T4 (1.0 g/kg) | T5 (2.0 g/kg) |
---|---|---|---|---|---|
Crude protein (%) | 62.82 ± 8.18 | 64.70 ± 6.09 | 61.52 ± 6.14 | 65.06 ± 9.01 | 69.41 ± 8.22 |
Crude lipid (%) | 4.14 ± 0.38 | 3.45 ± 0.23 | 3.87 ± 0.27 | 3.77 ± 0.03 | 3.68 ± 0.13 |
Moisture (%) | 3.35 ± 0.07 | 3.23 ± 0.04 | 3.72 ± 0.05 | 3.35 ± 0.00 | 3.13 ± 0.01 |
Ash (%) | 5.50 ± 0.25 | 5.59 ± 0.30 | 5.58 ± 0.11 | 4.64 ± 0.90 | 4.57 ± 0.30 |
Fillet (%) | 33.48 ± 2.14 | 34.16 ± 0.46 | 35.48 ± 11.04 | 33.07 ± 2.24 | 35.55 ± 4.74 |
Carcass (%) | 48.66 ± 2.70 | 46.12 ± 2.67 | 55.05 ± 2.13 | 50.56 ± 12.17 | 47.61 ± 4.91 |
Hepatosomatic index (HIS; %) | 2.42 ± 0.58 | 2.62 ± 0.72 | 2.78 ± 0.91 | 2.62 ± 0.55 | 2.71 ± 0.53 |
Viscerosomatic index (VSI; %) | 10.54 ± 1.77 | 10.91 ± 1.80 | 11.91 ± 3.57 | 10.47 ± 1.57 | 10.82 ± 1.32 |
Spleenosomatic index (SSI; %) | 0.20 ± 0.05 | 0.18 ± 0.050 | .25 ± 0.13 | 0.27 ± 0.03 | 0.21 ± 0.03 |
Parameter | T1 (0.0 g/kg) | T2 (0.1 g/kg) | T3 (0.2 g/kg) | T4 (1.0 g/kg) | T5 (2.0 g/kg) |
---|---|---|---|---|---|
Total protein (g/dL) | 2.66 ± 0.15 | 3.00 ± 0.26 | 2.70 ± 0.17 | 3.00 ± 0.36 | 2.90 ± 0.10 |
Albumin (g/dL) | 0.83 ± 0.05 | 0.80 ± 0.10 | 0.80 ± 0.02 | 0.90 ± 0.10 | 0.86 ± 0.05 |
Globulin (g/dL) | 1.83 ± 0.15 | 2.20 ± 0.17 | 1.90 ± 0.17 | 2.10 ± 0.26 | 2.03 ± 0.05 |
Aspartate aminotransferase; AST (U/L) | 73.66 ± 14.70 | 56.33 ± 16.66 | 69.33 ± 13.12 | 36.33 ± 15.30 | 54.66 ± 19.32 |
Alanine aminotransferase; ALT (U/L) | 39.33 ± 8.62 | 30.33 ± 16.01 | 48.33 ± 18.36 | 30.00 ± 10.15 | 55.66 ± 15.55 |
Blood Urea Nitrogen; BUN (mg/dL) | 1.33 ± 0.57 | 1.50 ± 0.50 | 1.16 ± 0.28 | 1.33 ± 0.29 | 1.35 ± 0.57 |
Cholesterol (mg/dL) | 123.66 ± 6.50 | 130.66 ± 10.21 | 117.33 ± 29.31 | 108.66 ± 21.34 | 134.00 ± 11.35 |
Parameter | T1 (0.0 g/kg) | T2 (0.1 g/kg) | T3 (0.2 g/kg) | T4 (1.0 g/kg) | T5 (2.0 g/kg) |
---|---|---|---|---|---|
Lysozyme activity (U/mL) | 5.80 ± 2.77 b | 7.30 ± 2.10 b | 11.00 ± 2.54 a | 11.52 ± 2.21 a | 12.16 ± 1.68 a |
Myeloperoxidase; MPO (OD450nm) | 2.00 ± 0.74 b | 1.76 ± 1.15 b | 2.59 ± 0.84 ab | 3.28 ± 0.17 a | 3.21 ± 0.19 a |
Superoxide dismutase; SOD (U/g liver) | 10.20 ± 1.48 c | 14.20 ± 0.83 b | 11.60 ± 3.64 bc | 16.60 ± 2.40 b | 20.80 ± 3.49 a |
Catalase activity; CAT (U/g liver) | 26.25 ± 9.27 c | 31.25 ± 9.88 bc | 39.06 ± 9.37 abc | 46.25 ± 11.35 a | 42.50 ± 5.22 ab |
Malondialdehyde; MDA (µmol/L) | 77.82 ± 8.90 a | 77.35 ± 6.75 a | 82.88 ± 20.42 a | 75.94 ± 7.92 a | 92.39 ± 10.10 a |
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Sutthi, N.; Wangkahart, E.; Panase, P.; Karirat, T.; Deeseenthum, S.; Ma, N.L.; Luang-In, V. Dietary Administration Effects of Exopolysaccharide Produced by Bacillus tequilensis PS21 Using Riceberry Broken Rice, and Soybean Meal on Growth Performance, Immunity, and Resistance to Streptococcus agalactiae of Nile tilapia (Oreochromis niloticus). Animals 2023, 13, 3262. https://doi.org/10.3390/ani13203262
Sutthi N, Wangkahart E, Panase P, Karirat T, Deeseenthum S, Ma NL, Luang-In V. Dietary Administration Effects of Exopolysaccharide Produced by Bacillus tequilensis PS21 Using Riceberry Broken Rice, and Soybean Meal on Growth Performance, Immunity, and Resistance to Streptococcus agalactiae of Nile tilapia (Oreochromis niloticus). Animals. 2023; 13(20):3262. https://doi.org/10.3390/ani13203262
Chicago/Turabian StyleSutthi, Nantaporn, Eakapol Wangkahart, Paiboon Panase, Thipphiya Karirat, Sirirat Deeseenthum, Nyuk Ling Ma, and Vijitra Luang-In. 2023. "Dietary Administration Effects of Exopolysaccharide Produced by Bacillus tequilensis PS21 Using Riceberry Broken Rice, and Soybean Meal on Growth Performance, Immunity, and Resistance to Streptococcus agalactiae of Nile tilapia (Oreochromis niloticus)" Animals 13, no. 20: 3262. https://doi.org/10.3390/ani13203262