Evaluation of Flathead Grey Mullets (Mugil cephalus) Immunization and Long-Term Protection against Vibrio harveyi Infection Using Three Different Vaccine Preparations
Abstract
:1. Introduction
2. Results
2.1. Fish Mortality Rates Post Challenges with Vibrio harveyi and Bacteriology Analysis
2.2. Production and Standardization of Mice Anti-Mullet Antibodies
2.3. ELISA Analysis of Mullet Anti-Vibrio harveyi Antibodies in Immunized Fish
2.4. ELISA Analysis to Detect Cross Reactivity against Other Vibrio spp.
2.5. Evaluation of Heath-Killed Bacteria’s Long-Term Protection, and Antibody Production
3. Discussion
4. Materials and Methods
4.1. Preparation of Three Different Vaccines
4.1.1. Isolation and Culture of Vibrio harveyi Strain RS2016
4.1.2. Heat-Killed Bacteria (HKB) Preparation
4.1.3. Denatured Membrane Proteins (BME) Preparation
4.1.4. Internal Proteins Preparation
4.2. Immunization and Challenge Infections
4.2.1. Fish
4.2.2. Experimental Design
4.2.3. Samples Collection and Analyses
4.3. Evaluation of Long-Term Protection and Antibody Production of the Heat-Killed Bacteria with Adjuvant Preparation
4.4. Production of Mice Anti Mullets Antibodies
4.4.1. Mice Immunization with Mullet Igs
4.4.2. Gel Electrophoresis and Western Blot Analysis
4.4.3. Dot–Blot Assay
4.5. Multilayer ELISA
4.5.1. Standardization of the Mice Anti-Mullet Serum for the ELISA
4.5.2. ELISA Analysis for Anti-Vibrio Harveyi Antibodies in Mullet Serum
4.5.3. ELISA Analysis to Detect Cross-Reactivity against Other Vibrio spp.
4.6. Statistical Analyses
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
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Chronology | Number of Bleedings | Time (Weeks) |
---|---|---|
Immunization | Bleeding Time 0 (T0) | 0 |
Bleeding Time 1 (T1) | 1 | |
Bleeding Time 2 (T2) | 2 | |
Booster | Bleeding Time 0 (T0B) | 4 |
Bleeding Time 1 (T1B) | 5 | |
Bleeding Time 2 (T2B) | 6 | |
First Challenge | Bleeding time 0 (T0C) | 7 |
Bleeding time 1 (T1C) | 8 | |
Bleeding time 2 (T2C) | 9 | |
Bleeding time 3 (T3C) | 10 | |
Bleeding time 4 (T4C) | 11 | |
Second Challenge | Bleeding time 5 (T5C) | 24 |
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Strem, R.; Meiri-Ashkenazi, I.; Segal, N.; Ehrlich, R.; Shashar, N.; Sharon, G. Evaluation of Flathead Grey Mullets (Mugil cephalus) Immunization and Long-Term Protection against Vibrio harveyi Infection Using Three Different Vaccine Preparations. Int. J. Mol. Sci. 2023, 24, 8277. https://doi.org/10.3390/ijms24098277
Strem R, Meiri-Ashkenazi I, Segal N, Ehrlich R, Shashar N, Sharon G. Evaluation of Flathead Grey Mullets (Mugil cephalus) Immunization and Long-Term Protection against Vibrio harveyi Infection Using Three Different Vaccine Preparations. International Journal of Molecular Sciences. 2023; 24(9):8277. https://doi.org/10.3390/ijms24098277
Chicago/Turabian StyleStrem, Rosa, Iris Meiri-Ashkenazi, Na’ama Segal, Roberto Ehrlich, Nadav Shashar, and Galit Sharon. 2023. "Evaluation of Flathead Grey Mullets (Mugil cephalus) Immunization and Long-Term Protection against Vibrio harveyi Infection Using Three Different Vaccine Preparations" International Journal of Molecular Sciences 24, no. 9: 8277. https://doi.org/10.3390/ijms24098277