title: Protective potential of recombinant non-purified botulinum neurotoxin serotypes C and D.
abstract: Botulinum neurotoxin (BoNT) serotypes C and D are responsible for cattle botulism, a fatal paralytic disease that results in great economic losses in livestock production. Vaccination is the main approach to prevent cattle botulism. However, production of commercially available vaccines (toxoids) involves high risk and presents variation of BoNT production between batches. Such limitations can be attenuated by the development of novel nontoxic recombinant vaccines through a simple and reproducible process. The aim of this study was to evaluate the protective potential of recombinant non-purified botulinum neurotoxin serotypes C and D. Bivalent vaccines containing 200 μg rHCC and rHCD each were formulated in three different ways: (1) purified antigens; (2) recombinant Escherichia coli bacterins; (3) recombinant E. coli cell lysates (supernatant and inclusion bodies). Guinea pigs immunized subcutaneously with recombinant formulations developed a protective immune response against the respective BoNTs as determined by a mouse neutralization bioassay with pooled sera. Purified recombinant antigens were capable of inducing 13 IU/mL antitoxin C and 21 IU/mL antitoxin D. Similarly, both the recombinant bacterins and the cell lysate formulations were capable of inducing 12 IU/mL antitoxin C and 20 IU/mL antitoxin D. These values are two times as high as compared to values induced by the commercial toxoid used as control, and two to ten times as high as the minimum amount required by the Brazilian Ministry of Agriculture, Livestock and Food Supply (MAPA), respectively. Therefore, we used a practical, industry-friendly, and efficient vaccine production process that resulted in formulations capable of inducing protective immune response (neutralizing antitoxins) against botulism serotypes C and D.
authors.affiliation: Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Clóvis
authors.initials: C
authors.lastname: Moreira
authors.affiliation: Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Carlos Eduardo Pouey
authors.initials: CE
authors.lastname: da Cunha
authors.affiliation: Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Gustavo Marçal Schmidt Garcia
authors.initials: GM
authors.lastname: Moreira
authors.affiliation: Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Mendonça
authors.affiliation: Faculdade de Medicina Veterinária, Universidade Federal do Pará, Castanhal, PA, Brazil.
authors.forename: Felipe Masiero
authors.initials: FM
authors.lastname: Salvarani
authors.affiliation: Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil; Faculdade de Nutrição, Universidade Federal de Pelotas, RS, Brazil.
authors.forename: Ângela Nunes
authors.initials: ÂN
authors.lastname: Moreira
authors.affiliation: Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil. Electronic address: fabricio.rochedo@ufpel.edu.br.
authors.forename: Fabricio Rochedo
authors.initials: FR
authors.lastname: Conceição
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000097:blood*; D000992:Antitoxins / Q000096:biosynthesis / Q000097:blood*; D001428:Bacterial Vaccines / Q000008:administration & dosage* / Q000096:biosynthesis / Q000276:immunology; D001905:Botulinum Toxins / Q000008:administration & dosage* / Q000096:biosynthesis / Q000276:immunology; D019274:Botulinum Toxins, Type A / Q000008:administration & dosage* / Q000096:biosynthesis / Q000276:immunology; D001906:Botulism / Q000097:blood / Q000276:immunology / Q000517:prevention & control*; D003014:Clostridium botulinum / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006168:Guinea Pigs; D056724:Immunity, Humoral / Q000187:drug effects; D051379:Mice; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic
chemical_list: D000907:Antibodies, Bacterial; D000992:Antitoxins; D001428:Bacterial Vaccines; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A; C096324:botulinum toxin type C
keywords: Cattle botulism; Cell lysate vaccine; Clostridium botulinum; Recombinant bacterins; Vaccine