Beatriz Almeida Sampaio (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Beatriz Almeida Sampaio Manoel Neres Santos Júnior (Microbiology Department, State University of Santa Cruz, Rod. Jorge Amado, Km a6, Ilhéus 45662-900, BA, Brazil) Manoel Neres Santos Júnior Bruna Carolina de Brito Guimarães (Microbiology Department, State University of Santa Cruz, Rod. Jorge Amado, Km a6, Ilhéus 45662-900, BA, Brazil) Bruna Carolina de Brito Guimarães Emilly Stefane Souza Andres (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Emilly Stefane Souza Andres Ágatha Morgana Bertoti da Silva (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Ágatha Morgana Bertoti da Silva Camila Pacheco Gomes (Microbiology Department, State University of Santa Cruz, Rod. Jorge Amado, Km a6, Ilhéus 45662-900, BA, Brazil) Camila Pacheco Gomes Rafaela de Souza Bittencourt (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Rafaela de Souza Bittencourt Thiago Macêdo Lopes Correia (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Thiago Macêdo Lopes Correia Lucas Santana Coelho da Silva (Microbiology Department, State University of Santa Cruz, Rod. Jorge Amado, Km a6, Ilhéus 45662-900, BA, Brazil) Lucas Santana Coelho da Silva Jurandir Ferreira da Cruz (Goat Sheep Sector, Department of Plant Science and Animal Science, State University of Southwest Bahia, Estrada do Bem Querer, Km 4, Vitória da Conquista 45083-900, BA, Brazil) Jurandir Ferreira da Cruz Rohini Chopra-Dewasthaly (Department of Pathobiology, Institute of Microbiology, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria) Rohini Chopra-Dewasthaly Guilherme Barreto Campos (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Guilherme Barreto Campos Jorge Timenetsky (Institute of Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes, 2415, Butantã 05508-900, SP, Brazil) Jorge Timenetsky Bruno Lopes Bastos (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Bruno Lopes Bastos Lucas Miranda Marques (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Lucas Miranda Marques Maysa Santos Barbosa (Institute of Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes, 2415, Butantã 05508-900, SP, Brazil) Maysa Santos Barbosa Matheus Gonçalves de Oliveira (Multidisciplinary Institute in Health, Federal University of Bahia, Rua Hormindo Barros 58, Vitória da Conquista 45029-094, BA, Brazil) Matheus Gonçalves de Oliveira MDPI 2025 Background/Objectives: Contagious agalactia (CA) is a disease typically caused by Mycoplasma agalactiae, affecting small ruminants worldwide and being endemic in certain countries. CA causes severe economic losses due to mastitis, agalactia, and arthritis. As an alternative to existing immunoprophylactic measures, this study aimed to develop a recombinant subunit vaccine against M. agalactiae and evaluate its specific immune response in goats. Methods: Goats were divided into three groups: group 1 received recombinant proteins (P40 and MAG_1560), group 2 received formalin-inactivated M. agalactiae, and group 3 received Tris-buffered saline (negative control). All solutions were emulsified in Freund’s adjuvant. Animals were monitored for 181 days. IgG antibody production was assessed by ELISA, and peripheral blood mononuclear cells (PBMCs) were analyzed by real-time PCR for the expression of IL-1β, IFN-γ, IL-12, and MHC class II genes. Results: M. agalactiae-specific antibody response was observed for six months in the sera of animals from group 1. Analysis of cytokine gene expression revealed increased IL-1β mRNA levels over time in both experimental groups. In group 1, IFN-γ mRNA levels increased with P40 stimulation and decreased with MAG_1560. IL-12 mRNA expression decreased over time in group 1 with P40 stimulation, whereas group 2 showed increased IL-12 expression for both proteins. MHC-II expression was stimulated in both groups. Conclusions: The recombinant proteins induced antibody production and cytokine expression, demonstrating immunogenic potential and supporting their promise as vaccine candidates capable of eliciting both humoral and cellular immune responses against M. agalactiae. PDFDocument eng 2026-02-11T09:28:11.345445Z 2025 Contagious Agalactia M. Agalactiae Recombinant; Subunit Vaccine 3928419 b application/pdf http://creativecommons.org/licenses/by/4.0/