Design of a New Vaccine Prototype against Porcine Circovirus Type 2 (PCV2), M. hyopneumoniae and M. hyorhinis Based on Multiple Antigens Microencapsulation with Sulfated Chitosan

Author:

Arrieta-Mendoza Darwuin1,Garces Bruno2,Hidalgo Alejandro A.2ORCID,Neira Victor3ORCID,Ramirez Galia3ORCID,Neira-Carrillo Andrónico4ORCID,Bucarey Sergio A.5ORCID

Affiliation:

1. Doctoral Program in Forestry, Agricultural and Veterinary Sciences, South Campus, University of Chile, Av. Santa Rosa 11315, La Pintana, Santiago 8820808, Chile

2. Escuela de Química y Farmacia, Facultad de Medicina, Universidad Andres Bello, 2320 Sazié, Santiago 8320000, Chile

3. Departamento de Medicina Preventiva, Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile, Av. Santa Rosa 11735, La Pintana, Santiago 8320000, Chile

4. Laboratorio Polyforms, Departamento de Ciencias Biológicas, Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile, Av. Santa Rosa 11735, La Pintana, Santiago 8320000, Chile

5. Centro Biotecnológico Veterinario, Biovetec, Departamento de Ciencias Biológicas, Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile, Av. Santa Rosa 11735, La Pintana, Santiago 8320000, Chile

Abstract

This work evaluated in vivo an experimental-multivalent-vaccine (EMV) based on three Porcine Respiratory Complex (PRC)-associated antigens: Porcine Circovirus Type 2 (PCV2), M. hyopneumoniae (Mhyop) and M. hyorhinis (Mhyor), microencapsulated with sulfated chitosan (M- ChS + PRC-antigens), postulating chitosan sulphate (ChS) as a mimetic of the heparan sulfate receptor used by these pathogens for cell invasion. The EMV was evaluated physicochemically by SEM (Scanning-Electron-Microscopy), EDS (Energy-Dispersive-Spectroscopy), Pdi (Polydispersity-Index) and zeta potential. Twenty weaned pigs, distributed in four groups, were evaluated for 12 weeks. The groups 1 through 4 were as follows: 1-EMV intramuscular-route (IM), 2-EMV oral-nasal-route (O/N), 3-Placebo O/N (M-ChS without antigens), 4-Commercial-vaccine PCV2-Mhyop. qPCR was used to evaluate viral/bacterial load from serum, nasal and bronchial swab and from inguinal lymphoid samples. Specific humoral immunity was evaluated by ELISA. M-ChS + PRC-antigens measured between 1.3–10 μm and presented low Pdi and negative zeta potential, probably due to S (4.26%). Importantly, the 1-EMV protected 90% of challenged animals against PCV2 and Mhyop and 100% against Mhyor. A significant increase in antibody was observed for Mhyor (1-EMV and 2-EMV) and Mhyop (2-EMV), compared with 4-Commercial-vaccine. No difference in antibody levels between 1-EMV and 4-Commercial-vaccine for PCV2-Mhyop was observed. Conclusion: The results demonstrated the effectiveness of the first EMV with M-ChS + PRC-antigens in pigs, which were challenged with Mhyor, PCV2 and Mhyop, evidencing high protection for Mhyor, which has no commercial vaccine available.

Funder

Chilean Agency for Research and Development (ANID) Fondef Idea I + D project

Bucarey and Fondo de Investigación Veterinaria de la Facultad de Cs. Veterinarias y Pecuarias de la Universidad de Chile (FIV2022) project

program Doctorate in Forestry, Agricultural and Veterinary Sciences, University of Chile

Publisher

MDPI AG

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