Vaccine Efficacy of a Newly Developed Feed-Based Whole-Cell Polyvalent Vaccine against Vibriosis, Streptococcosis and Motile Aeromonad Septicemia in Asian Seabass, Lates calcarifer
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains and Culture Conditions
2.2. Reviving the Virulence and Preparation of Pathogenic Bacterial Strains
PCR Amplification Using 16S rRNA
2.3. Feed-Based Vaccine Preparation
2.4. Feed Quality Analysis
2.4.1. Proximate Analysis
2.4.2. Physical Stability of Feed in Water
2.4.3. Palatability Test
2.4.4. Growth Performance
2.5. Vaccination Trial
2.5.1. The Fish
2.5.2. Experimental Design
2.5.3. Challenge Test
2.6. Sample Processing
2.6.1. Enzyme-Linked Immunosorbent Assay (ELISA)
2.6.2. Serum Lysozyme Activity
2.6.3. Quantitative Real-Time PCR (qPCR)
2.7. Statistical Analysis
3. Results
3.1. Feed Quality Analysis and Growth Performance
3.2. IgM Antibody Responses
3.3. Lysozyme Activity
3.4. Gene Expression
3.5. Efficacy of the Feed-Based Polyvalent Vaccine
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Bacterial Strain | Description/Genotype | Source | Ref. |
---|---|---|---|
Pathogenic V. harveyi strain Vh1 | Wild type, local isolate | Lab collection | [32] |
Pathogenic A. hydrophila strain Ah1sa5 | Wild type, local isolate | Lab collection | [33] |
Pathogenic S. agalactiae strain Sa2k | Wild type, local isolate | Lab collection | [30] |
Primers | Primer Sequence (5’-3’) | Tm (°C) | Expected Size (bp) | Reference |
---|---|---|---|---|
16S rRNA_R | AGAGTTTGATCCTGGCTCAG | 54.0 | 1541 | [34] |
16S rRNA_F | GGTTACCTTGTTACGACTT |
Target | Sequences (5’-3’) | Product Size | Annealing Temperature (˚C) | References | Note |
---|---|---|---|---|---|
Dendritic cells | F:AACAGCACACGCTCACTCAC R:CGATCATGTGAGCCTTGAGA | 153 | 60 | Zoccola et al. [50] | Initiate an adaptive immune response |
C3 | F:GCAATCCTCCACAACTACAG R:ACTCTGACCTCCTGACGATAC | 93 | 59 | Mohd-Shaharuddin et al. [51] | Innate defense against common pathogens |
CCL4 | F:TCCTCGTCTCACTCTGTCTGT R:GACCTGCCACTGTCTTCAGC | 301 | 60 | Chin et al. [36] | Chemokine attracts innate immune cells |
MHC I | F:GGCTGTTTTTGCCGCTCTG R:GTGGACAGGTCTGGATAAAG | 112 | 60 | Chin et al. [36] | Molecule-presenting antigen for CD8+ |
β-actin (control) | F:TACCACCGGTATCGTCATGGA R:CCACGCTCTGTCAGGATCTTC | 126 | 60 | Chin et al. [36] | Reference gene |
Composition | Feed | |
---|---|---|
Control | Vaccinated | |
Crude protein (%) | 32 ± 7.2 | 33.1 ± 6.0 |
Crude lipid (%) | 3 ± 1.0 | 4 ± 0.5 |
Crude fiber (%) | 6.8 ± 2 | 6.2 ± 0.0 |
Ash (%) | 11.6 ± 1.8 | 10.8 ± 2.11 |
Moisture (%) | 12 ± 3.3 | 13 ± 1.4 * |
Carbohydrate (%) | 34.6 ± 7.65 | 32.9 ± 5.0 |
Parameters | Treatment Group | |
---|---|---|
Control Group | Vaccinated Group | |
Initial body weight (g) | 2.31 ± 0.08 | 2.31 ± 0.08 |
Final body weight (g) | 9.78 ± 0.05 | 17.5 ± 2.23 * |
Weight gain (g) | 7.63 ± 1.11 | 14.35 ± 1.09 * |
SGR (%/day) | 0.36 ± 0.03 | 0.71 ± 0.05 * |
FCR (g/g) | 0.28 ± 0.06 | 0.16 ± 0.01 * |
Group | Number of Fish | Primary Vaccination (Day 0–5, Oral, 5% Fish Body Weight) | Booster Dose (Day 14–18, Oral, 5% Fish Body Weight | Challenge Group (10 Fish/Tank in Duplicates) | Challenge Dose/Fish (Day 28, IP, 0.1 mL/Fish) | RPS (%) |
---|---|---|---|---|---|---|
Control | 80 | PBS + POA | PBS + POA | Control (+PBS) | PBS | - |
Control (+Vh) | 107 CFU Vh | - | ||||
Control (+Sa) | 107 CFU Sa | - | ||||
Control (+Ah) | 107 CFU Ah | - | ||||
Vaccinated | 80 | 106 cells/kg of feed + POA | 106 cells/kg of feed + POA | Vaccinated (+PBS) | PBS | - |
Vaccinated (+Vh) | 107 CFU Vh | 75 ± 7.07 | ||||
Vaccinated (+Sa) | 107 CFU Sa | 80 ± 0 | ||||
Vaccinated (+Ah) | 107 CFU Ah | 80 ± 0 |
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Mohamad, A.; Zamri-Saad, M.; Amal, M.N.A.; Al-saari, N.; Monir, M.S.; Chin, Y.K.; Md Yasin, I.-S. Vaccine Efficacy of a Newly Developed Feed-Based Whole-Cell Polyvalent Vaccine against Vibriosis, Streptococcosis and Motile Aeromonad Septicemia in Asian Seabass, Lates calcarifer. Vaccines 2021, 9, 368. https://doi.org/10.3390/vaccines9040368
Mohamad A, Zamri-Saad M, Amal MNA, Al-saari N, Monir MS, Chin YK, Md Yasin I-S. Vaccine Efficacy of a Newly Developed Feed-Based Whole-Cell Polyvalent Vaccine against Vibriosis, Streptococcosis and Motile Aeromonad Septicemia in Asian Seabass, Lates calcarifer. Vaccines. 2021; 9(4):368. https://doi.org/10.3390/vaccines9040368
Chicago/Turabian StyleMohamad, Aslah, Mohd Zamri-Saad, Mohammad Noor Azmai Amal, Nurhidayu Al-saari, Md. Shirajum Monir, Yong Kit Chin, and Ina-Salwany Md Yasin. 2021. "Vaccine Efficacy of a Newly Developed Feed-Based Whole-Cell Polyvalent Vaccine against Vibriosis, Streptococcosis and Motile Aeromonad Septicemia in Asian Seabass, Lates calcarifer" Vaccines 9, no. 4: 368. https://doi.org/10.3390/vaccines9040368