title: Anti-botulism single-shot vaccine using chitosan for protein encapsulation by simple coacervation.
abstract: The aim of the present study was to compare the potency and safety of vaccines against Clostridium botulinum (C. botulinum) type C and D formulated with chitosan as controlled release matrix and vaccines formulated in conventional manner using aluminum hydroxide. Parameters were established for the development of chitosan microspheres, using simple coacervation to standardize the use of this polymer in protein encapsulation for vaccine formulation. To formulate a single shot vaccine inactivated antigens of C. botulinum type C and D were used with original toxin titles equal to 5.2 and 6.2 log LD50/ml, respectively. For each antigen a chitosan based solution of 50 mL was prepared. Control vaccines were formulated by mixing toxoid type C and D with aluminum hydroxide [25% Al(OH)3, pH 6.3]. The toxoid sterility, innocuity and potency of vaccines were evaluated as stipulated by MAPA-BRASIL according to ministerial directive no. 23. Encapsulation efficiency of BSA in chitosan was 32.5-40.37%, while that the encapsulation efficiency to toxoid type C was 41,03% (1.94 mg/mL) and of the toxoid type D was 32.30% (1.82 mg/mL). The single shot vaccine formulated using chitosan for protein encapsulation through simple coacervation showed potency and safety similar to conventional vaccine currently used in Brazilian livestock (10 and 2 IU/mL against C. botulinum type C and D, respectively). The present work suggests that our single shot vaccine would be a good option as a cattle vaccine against these C. botulinum type C and D.
authors.affiliation: Vallée S.A., Av. Comendador António Loureiro Ramos, 1500, Cidade Industrial, 39404-003 Montes Claros, MG, Brazil. Electronic address: rogershoji@gmail.com.
authors.forename: Roger S
authors.initials: RS
authors.lastname: Sari
authors.affiliation: Instituto de Ciéncias Agrárias, Universidade Federal de Minas Gerais, Av. Universitária, 1000, Universitário, 39404-547 Montes Claros, MG, Brazil. Electronic address: annachristinadealmeida@gmail.com.
authors.forename: Anna Christina
authors.initials: AC
authors.lastname: de Almeida
authors.affiliation: Engenharia de Bioprocessos e Biotecnologia, Universidade Federal do Tocantins, Gurupi, TO, Brazil. Electronic address: alexcangussu@uft.edu.br.
authors.forename: Alex S R
authors.initials: ASR
authors.lastname: Cangussu
authors.affiliation: Vallée S.A., Av. Comendador António Loureiro Ramos, 1500, Cidade Industrial, 39404-003 Montes Claros, MG, Brazil. Electronic address: edvander@hotmail.com.
authors.forename: Edson V
authors.initials: EV
authors.lastname: Jorge
authors.affiliation: Vallée S.A., Av. Comendador António Loureiro Ramos, 1500, Cidade Industrial, 39404-003 Montes Claros, MG, Brazil.
authors.forename: Otto D
authors.initials: OD
authors.lastname: Mozzer
authors.affiliation: Instituto Federal do Norte de Minas Gerais Campus Araçuaí, Rodovia. BR 367, km 278, Itaobim/Araçuaí, 39600-000 Araçuaí, MG, Brasil. Electronic address: htacilio@hotmail.com.
authors.forename: Hércules Otacílio
authors.initials: HO
authors.lastname: Santos
authors.affiliation: Controle de Qualidade, Instituto do Butantan, Avenida Vital Brasil, 1500, Butantan, 05503-900 Sáo Paulo, SP, Brazil. Electronic address: wagner.quintilio@butantan.gov.br.
authors.forename: Wagner
authors.initials: W
authors.lastname: Quintilio
authors.affiliation: Instituto de Ciéncias Agrárias, Universidade Federal de Minas Gerais, Av. Universitária, 1000, Universitário, 39404-547 Montes Claros, MG, Brazil. Electronic address: ibrandi@hotmail.com.
authors.forename: Igor Viana
authors.initials: IV
authors.lastname: Brandi
authors.affiliation: Instituto de Ciéncias Agrárias, Universidade Federal de Minas Gerais, Av. Universitária, 1000, Universitário, 39404-547 Montes Claros, MG, Brazil. Electronic address: andrade_viviane@hotmail.com.
authors.forename: Viviane Aguiar
authors.initials: VA
authors.lastname: Andrade
authors.affiliation: Departamento Tecnologia Farmacéutica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. Electronic address: samir@pharma.ufrj.br.
authors.forename: Angelo Samir M
authors.initials: ASM
authors.lastname: Miguel
authors.affiliation: Instituto Federal do Norte de Minas Gerais Campus Araçuaí, Rodovia. BR 367, km 278, Itaobim/Araçuaí, 39600-000 Araçuaí, MG, Brasil. Electronic address: elianemsobrinho@hotmail.com.
authors.forename: Eliane M
authors.initials: EM
authors.lastname: Sobrinho Santos
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001428:Bacterial Vaccines / Q000008:administration & dosage* / Q000009:adverse effects / Q000737:chemistry / Q000276:immunology; D001906:Botulism / Q000517:prevention & control*; D048271:Chitosan; D003014:Clostridium botulinum / Q000276:immunology; D003692:Delayed-Action Preparations; D006168:Guinea Pigs; D014677:Pharmaceutical Vehicles; D064166:Vaccine Potency
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D003692:Delayed-Action Preparations; D014677:Pharmaceutical Vehicles; D048271:Chitosan
keywords: Chitosan; Controlled antigen release; Neurotoxins; Simple coacervation; Single-shot vaccine