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Home / Equine Research Bank / PLoS One / Characterization of clinically-attenuated Burkholderia malle...
PloS one2008; 3(4); e2058; doi: 10.1371/journal.pone.0002058

Characterization of clinically-attenuated Burkholderia mallei by whole genome sequencing: candidate strain for exclusion from Select Agent lists.

Abstract: Burkholderia mallei is an understudied biothreat agent responsible for glanders which can be lethal in humans and animals. Research with this pathogen has been hampered in part by constraints of Select Agent regulations for safety reasons. Whole genomic sequencing (WGS) is an apt approach to characterize newly discovered or poorly understood microbial pathogens. Results: We performed WGS on a strain of B. mallei, SAVP1, previously pathogenic, that was experimentally infected in 6 equids (4 ponies, 1 mule, 1 donkey), natural hosts, for purposes of producing antibodies. Multiple high inocula were used in some cases. Unexpectedly SAVP1 appeared to be avirulent in the ponies and mule, and attenuated in the donkey, but induced antibodies. We determined the genome sequence of SAVP1 and compared it to a strain that was virulent in horses and a human. In comparison, this phenotypic avirulent SAVP1 strain was missing multiple genes including all the animal type III secretory system (T3SS) complex of genes demonstrated to be essential for virulence in mice and hamster models. The loss of these genes in the SAVP1 strain appears to be the consequence of a multiple gene deletion across insertion sequence (IS) elements in the B. mallei genome. Therefore, the strain by itself is unlikely to revert naturally to its virulent phenotype. There were other genes present in one strain and not the other and vice-versa. Conclusions: The discovery that this strain of B. mallei was both avirulent in the natural host ponies, and did not possess T3SS associated genes may be fortuitous to advance biodefense research. The deleted virulence-essential T3SS is not likely to be re-acquired naturally. These findings may provide a basis for exclusion of SAVP1 from the Select Agent regulation or at least discussion of what else would be required for exclusion. This exclusion could accelerate research by investigators not possessing BSL-3 facilities and facilitate the production of reagents such as antibodies without the restraints of Select Agent regulation.
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Publication Date: 2008-04-30 PubMed ID: 18446194PubMed Central: PMC2312325DOI: 10.1371/journal.pone.0002058Google Scholar: Lookup
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Summary

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The research describes the application of whole genomic sequencing to understand the characteristics and virulence of Burkholderia mallei, a substantial biothreat agent. The genomic changes of an attenuated strain known as SAVP1, which lost its virulent qualities after experimental infection in various equids, were particularly analyzed.

Methodology of Research

  • The team used Whole Genomic Sequencing (WGS) on a particular strain of Burkholderia mallei known as SAVP1. This strain was formerly pathogenic and was artificially infected in various equids such as ponies and mules to stimulate antibody production.
  • Contrary to the expectations, the SAVP1 strain demonstrated avirulent behavior in ponies and mules, and attenuated responses in donkeys, while still succeeding in inducing antibodies.

Genomic Comparison and Findings

  • The researchers then sequenced the genome of SAVP1 and compared it to a strain that had shown virulence in humans and horses.
  • The comparison revealed that the SAVP1 strain was lacking several essential genes, including the animal type III secretory system (T3SS) complex of genes that are fundamental for virulence in mouse and hamster models. The loss of these genes seems to be due to a multi-gene deletion across Insertion Sequence (IS) elements in the genome of Burkholderia mallei.
  • The researchers concluded that this deletion makes it highly unlikely for the strain to naturally revert to its virulent state.
  • Apart from the absence of T3SS genes, there were also other genes present in one strain and absent in the other, and vice versa.

Significance of the Findings

  • The underwhelming virulence of the SAVP1 strain and its lack of T3SS associated genes are useful discoveries that may aid in advancing biodefense research.
  • The absence of the T3SS, taken together with its unlikely natural re-acquisition, may justify excluding the SAVP1 strain from Select Agent regulations. This exclusion could encourage rapid research progress by investigators who lack BSL-3 facilities.
  • Further, this regulation could alleviate the restrictions associated with producing reagents such as antibodies, thereby facilitating research and response protocols against biological threats.

Cite This Article

APA
Schutzer SE, Schlater LR, Ronning CM, DeShazer D, Luft BJ, Dunn JJ, Ravel J, Fraser-Liggett CM, Nierman WC. (2008). Characterization of clinically-attenuated Burkholderia mallei by whole genome sequencing: candidate strain for exclusion from Select Agent lists. PLoS One, 3(4), e2058. https://doi.org/10.1371/journal.pone.0002058

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 3
Issue: 4
Pages: e2058
PII: e2058

Researcher Affiliations

Schutzer, Steven E
  • Department of Medicine, University of Medicine and Dentistry-New Jersey Medical School, Newark, New Jersey, United States of America. schutzer@umdnj.edu
Schlater, Linda R K
    Ronning, Catherine M
      DeShazer, David
        Luft, Benjamin J
          Dunn, John J
            Ravel, Jacques
              Fraser-Liggett, Claire M
                Nierman, William C

                  MeSH Terms

                  • Animals
                  • Bacterial Proteins / genetics
                  • Burkholderia mallei / genetics
                  • Burkholderia mallei / pathogenicity
                  • Chromosomes, Bacterial
                  • Equidae / microbiology
                  • Genes, Bacterial
                  • Genome, Bacterial / genetics
                  • Phenotype
                  • Sequence Analysis, DNA / methods
                  • Software
                  • Virulence

                  Grant Funding

                  • N01-AI30071 / NIAID NIH HHS
                  • N01AI30071 / NIAID NIH HHS
                  • AI063757 / NIAID NIH HHS
                  • U01 AI056480 / NIAID NIH HHS
                  • R43 AI063757 / NIAID NIH HHS

                  Conflict of Interest Statement

                  The authors have declared that no competing interests exist.

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