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Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis.

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  • Additional Information
    • Author(s):
    • Address:
      Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, São Paulo, Brazil.
    • Publication Information:
      Brazil
    • Abstract:
      Canine brucellosis is a zoonotic disease, caused by Brucella canis and induces reproductive failures. This study had the objective of investigate a B. canis outbreak in a breeding kennel, describe a multistep approach to characterize B. canis isolates and identify B. canis-specific proteins reacting with antibodies from naturally infected dogs. The kennel was located in São Paulo-SP, Brazil. At the time of sampling, in 2014, the kennel comprised 17 adult Beagle dogs. Blood samples were used both to isolate the bacteria and to detect the Brucella sp. DNA by PCR. Sera samples were used to detect antibodies reaction against B. canis using immunocromatography, rapid slide agglutination test with or without 2-mercaptoethanol and two sorts of ELISA kit. The isolates were characterized by phenotyping and genotyping methods and by mass spectrometry. The proteins from the isolates were separated using one and two dimension polyacrylamide gel electrophoresis (SDS PAGE/2DE), then they were submitted to sera from bacteremic, non-bacteremic or non-B. canis infected dogs using western immunoblotting. The reacted proteins spots were identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). Vaginal discharge, abortion, stillbirth, conception failure and general lymphopathy were found in the infected dogs. 24 Gram-negative, coccoid rod-shaped bacteria were isolated. Antibodies reactions against B. canis were observed at least once by the performed serological tests. Mass spectrometry analysis assigned all isolates to the genus Brucella sp. The phenotypic data clearly identified B. canis with only slight differences in phage typing patterns. The genomic data proved a single outbreak strain exhibiting only seven single-nucleotide polymorphisms in six isolates. SDS PAGE/2DE separated protein spots were identified by MALDI-TOF MS as 93 different proteins, out of them, 21 were identified in infected dogs (bacteremic and non-bacteremic phase of the infection) and were not identified in non-Brucella canis infected dogs. These proteins are candidates to be applied as antigens for the serodiagnosis improvement. A multistep approach involving classical microbiological methods, mass spectrometry and genomic sequencing can help us to characterize Brucella canis in naturally infected dogs and allows outbreak investigations. A reliable diagnosis is important to avoid economic losses in kennels and to prevent infection in dogs as well as the transmission toward humans.
    • Number of References:
      227 ref.
    • Subject Terms:
      Veterinary Science;Veterinary Science
    • Subject Terms:
    • Accession Number:
      antigenicity, bacterial infections, bacterioses, bacterium, enzyme linked immunosorbent assay, immunogens, isolates, matrix-associated laser desorption ionization time of flight mass spectrometry, PCR, serological diagnosis, undulant fever, zoonotic infections
    • CABICODES:
      Host Resistance and Immunity (HH600)
      Pets and Companion Animals (LL070)
      Prion, Viral, Bacterial and Fungal Pathogens of Animals (LL821) (New March 2000)
      Diagnosis of Animal Diseases (LL886) (New March 2000)
      General Molecular Biology (ZZ360) (Discontinued March 2000)
      Taxonomy and Evolution (ZZ380)
      Genetics and Molecular Biology of Microorganisms (ZZ395) (New March 2000)
      Techniques and Methodology (ZZ900)
    • Publication Information:
      Thesis
    • Accession Number:
      20193280074
    • Copyright:
      ©2019 CAB International
  • Citations
    • ABNT:
      SILVA, A. V. da. Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis. 2018. Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, São Paulo; Brazil, 2018. Disponível em: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074. Acesso em: 26 nov. 2020.
    • AMA:
      Silva AV da. Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis. Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis. 2018:90. Accessed November 26, 2020. http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074
    • APA:
      Silva, A. V. da. (2018). Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis [Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia]. In Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis (p. 90).
    • Chicago/Turabian: Author-Date:
      Silva, A. V. da. 2018. “Immunoproteomic Characterization of Brucella Canis to Identify Proteins Candidates as Antigens for Serodiagnosis.” Immunoproteomic Characterization of Brucella Canis to Identify Proteins Candidates as Antigens for Serodiagnosis. São Paulo; Brazil: Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia. http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074.
    • Harvard:
      Silva, A. V. da (2018) Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis, Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis. Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia. Available at: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074 (Accessed: 26 November 2020).
    • Harvard: Australian:
      Silva, AV da 2018, ‘Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis’, Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, São Paulo; Brazil, Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis, p. 90, viewed 26 November 2020, .
    • MLA:
      Silva, A. V.da. “Immunoproteomic Characterization of Brucella Canis to Identify Proteins Candidates as Antigens for Serodiagnosis.” Immunoproteomic Characterization of Brucella Canis to Identify Proteins Candidates as Antigens for Serodiagnosis, Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, 2018, p. 90. EBSCOhost, search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074.
    • Chicago/Turabian: Humanities:
      Silva, A. V. da. “Immunoproteomic Characterization of Brucella Canis to Identify Proteins Candidates as Antigens for Serodiagnosis.” Immunoproteomic Characterization of Brucella Canis to Identify Proteins Candidates as Antigens for Serodiagnosis. Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, 2018. http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074.
    • Vancouver/ICMJE:
      Silva AV da. Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis [Internet]. Immunoproteomic characterization of Brucella canis to identify proteins candidates as antigens for serodiagnosis. [São Paulo; Brazil]: Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia; 2018 [cited 2020 Nov 26]. p. 90. Available from: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=lhh&AN=20193280074