# Effectiveness of a Bivalent Recombinant Vaccine on the Production of Neutralizing Antibodies Against BoNT/C, BoNT/D, BoNT/CD e BoNT/DC in Bovines

## TL;DR

A bivalent recombinant vaccine was tested in cattle for its ability to produce antibodies against botulinum toxins C and D, showing strong effectiveness but limited protection against a toxin variant.

## Contribution

The study introduces a bivalent recombinant vaccine targeting BoNT/C and BoNT/D and compares its efficacy to a commercial toxoid vaccine in cattle.

## Key findings

- The recombinant vaccine significantly increased IgG antibody levels against BoNT/C and BoNT/D in vaccinated calves.
- Neutralizing antibody titers against BoNT/C and BoNT/D were higher in the recombinant vaccine group than in the toxoid vaccine group.
- The vaccine showed lower neutralizing activity against the mosaic variant BoNT/DC compared to the toxoid vaccine.

## Abstract

Background/Objectives. Bovine botulism, although relatively rare, presents significant economic losses due to high mortality rates and restrictions on livestock product trade. Vaccination remains the most effective strategy for preventing botulism-related mortality. This study evaluated the efficacy of a bivalent recombinant vaccine targeting the C-terminal portion of the heavy chain (Hc) of botulinum neurotoxin serotype C (BoNT/C) (Hc BoNT/C) and botulinum neurotoxin serotype D (BoNT/D) (Hc BoNT/D) in inducing neutralizing antibodies against these toxins and their mosaic variants BoNT/CD and BoNT/DC in cattle. This comparison aims to improve the design of an optimal recombinant vaccine for preventing bovine botulism caused by the most common serotypes. Methods. Twenty, four-month-old Holstein Friesian calves were randomly assigned to two groups of ten animals: vaccinated group and control group. Sera were collected at various time points to assess antibody titers using ELISA and neutralizing antibody titers using a mouse protection assay. Neutralizing antibody titers were compared to those obtained with a commercially available toxoid vaccine. Results. The recombinant vaccine elicited significant increases in anti-HcBoNT/C and anti-HcBoNT/D IgG antibody levels in vaccinated animals compared to controls animals with no adverse effects. Specifically, post-vaccination, the calves showed no local reactions (swelling, warmth) or behavioral changes suggestive of systemic illness. Neutralizing antibody titers against BoNT/C and BoNT/D were significantly higher in the recombinant vaccine group compared to the toxoid vaccine group. However, the recombinant vaccine showed lower neutralizing activity against BoNT/DC compared to the toxoid vaccine. Conclusions. The bivalent recombinant vaccine demonstrated promising immunogenicity in cattle, inducing high neutralizing antibody titers against BoNT/C and BoNT/D. While effective against these toxins, the lower efficacy against BoNT/DC highlights the need for further research to optimize the vaccine formulation, potentially by incorporating a BoNT/DC Hc component, to provide broader protection against bovine botulism.