Protective Immunization of Atlantic Salmon (Salmo salar L.) against Salmon Lice (Lepeophtheirus salmonis) Infestation
Abstract
:1. Introduction
2. Material and Methods
2.1. Experimental Facility and Ethics
2.2. Vaccine Preparation
2.3. Fish and Design of the Trial
2.4. Lice Challenge Procedures
2.5. Lice Counts
2.6. Fish Sampling
2.7. RNA Isolation and cDNA Synthesis
2.8. RT-qPCR Protocol
2.9. Enzyme-Linked Immunosorbent Assay (ELISA)
2.10. Immunohistochemistry (IHC)
2.10.1. Analysis of Total IgM and Production of IgM Positive Cells
2.10.2. Analysis of P33-Specific Antibodies In Situ in Spleen
3. Results
3.1. P33 Vaccination Reduces Salmon Lice Counts on Atlantic Salmon
3.2. P33 Vaccination Induces P33-Specific IgM Antibodies
3.2.1. Quantification of P33-Specific IgM in Plasma
3.2.2. Demonstration of P33-Specific Antibodies In Situ in the Spleen of Vaccinated Fish
3.3. Immunoglobulin Kinetics Are Altered after P33 Vaccination
3.3.1. P33 Vaccine Modifies Immunoglobulin Gene Expression in Vaccinated Fish
3.3.2. IgM Production in Fish Immune Tissues Is Modulated after P33 Vaccination
3.4. Vaccine-Induced Protection against Salmon Lice Correlates with the P33-Specific IgM in Fish Plasma
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Gene Name | Primers Sequences (5′–3′) | Accession Number | Effeciency (%) |
---|---|---|---|
Secretory immunoglobulin M (sIgM) | F:CTACAAGAGGGAGACCGGAG R:AGGGTCACCGTATTATCACTAGTT | XM_014203125 | 100.36 |
Membrane immunoglobulin M (mIgM) | F:CCTACAAGAGGGAGACCGA R:GATGAAGGTGAAGGCTGTTTT | Y12457 | 81.84 |
Immunoglobulin T isoform C (IgT-C) | F:GCTAAGAGTGTCTGGGAAATGA R:TGGAGGGTTTGAGATTGGTC | HQ379938.1 | 84.9 |
Immunoglobulin D (IgD) | F:TGAACATCGCTGCTTCAAC R:CCAGCACAGCACTGTCTCC | AF141606.1 | 97.09 |
Secretory immunoglobulin T isoform A (sIgT-A) | F:CCAAGGATAAGTGGGAGAGAA R:TCACTTGTCTTCACATGAGTTACC | GQ907003 | 80.2 |
Membrane immunoglobulin T isoform A (mIgT-A) | F:CCAAGGATAAGTGGGAGAGAA R:AGGATGTTCGCCATGGACT | GQ907003 | 103.41 |
Secretory immunoglobulin T isoform B (sIgT-B) | F:GAATGTTTGGGACACGGAAG R:TCACATATCTTGACATGAGTTACCC | GQ907004.1 | 89.89 |
Membrane immunoglobulin T isoform B (mIgT-B) | F:GAATGTTTGGGACACGGAAG R:GCTCAGTCAGTGGGATGTTCT | GQ907004.1 | 92.3 |
Elongation factor-1alpha (EF-1 α) | F:TGCCCCTCCAGGATGTCTAC R:CACGGCCCACAGGTACTG | BG933897 | 106.98 |
Tissue | Scoring Scale | Score Description |
---|---|---|
Spleen and anterior kidney | Abundance of IgM+ cells/microscopic field (10× magnification objective lens) | |
0 | No positive cells | |
1 | ≤10 cells | |
2 | 30–50 cells | |
3 | 60–80 cells | |
4 | >90 cells | |
IgM labeling intensity | ||
0 | No labeling | |
1 | Mild labeling | |
2 | Moderate labeling | |
3 | Strong labeling | |
4 | Heavy labeling |
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Tartor, H.; Karlsen, M.; Skern-Mauritzen, R.; Monjane, A.L.; Press, C.M.; Wiik-Nielsen, C.; Olsen, R.H.; Leknes, L.M.; Yttredal, K.; Brudeseth, B.E.; et al. Protective Immunization of Atlantic Salmon (Salmo salar L.) against Salmon Lice (Lepeophtheirus salmonis) Infestation. Vaccines 2022, 10, 16. https://doi.org/10.3390/vaccines10010016
Tartor H, Karlsen M, Skern-Mauritzen R, Monjane AL, Press CM, Wiik-Nielsen C, Olsen RH, Leknes LM, Yttredal K, Brudeseth BE, et al. Protective Immunization of Atlantic Salmon (Salmo salar L.) against Salmon Lice (Lepeophtheirus salmonis) Infestation. Vaccines. 2022; 10(1):16. https://doi.org/10.3390/vaccines10010016
Chicago/Turabian StyleTartor, Haitham, Marius Karlsen, Rasmus Skern-Mauritzen, Adérito Luis Monjane, Charles McLean Press, Christer Wiik-Nielsen, Rolf Hetlelid Olsen, Lisa Marie Leknes, Karine Yttredal, Bjørn Erik Brudeseth, and et al. 2022. "Protective Immunization of Atlantic Salmon (Salmo salar L.) against Salmon Lice (Lepeophtheirus salmonis) Infestation" Vaccines 10, no. 1: 16. https://doi.org/10.3390/vaccines10010016