Prevalence of Bacillus anthracis-like organisms and bacteriophages in the intestinal tract of the earthworm Eisenia fetida

Appl Environ Microbiol. 2010 Apr;76(7):2286-94. doi: 10.1128/AEM.02518-09. Epub 2010 Jan 29.

Abstract

Stable infection of Bacillus anthracis laboratory strains with environmental bacteriophages confers survival phenotypes in soil and earthworm intestinal niches (R. Schuch and V. A. Fischetti, PLoS One 4:e6532, 2009). Here, the natural occurrence of two such B. anthracis-infective bacteriophages, Wip1 and Wip4, was examined in the intestines of Eisenia fetida earthworms as part of a 6-year longitudinal study at a Pennsylvania forest site. The Wip1 tectivirus was initially dominant before being supplanted by the Wip4 siphovirus, which was then dominant for the next 3 years. In a host range analysis of a wide-ranging group of Bacillus species and related organisms, Wip1 and Wip4 were both infective only toward B. anthracis and certain B. cereus strains. The natural host of Wip4 remained constant for 3 years and was a B. cereus strain that expressed a B. anthracis-like surface polysaccharide at septal positions on the cell surface. Next, a novel metagenomic approach was used to determine the extent to which such B. cereus- and B. anthracis-like strains are found in worms from two geographical locations. Three different enrichment strategies were used for metagenomic DNA isolation, based either on the ability of B. cereus sensu lato to form heat-resistant spores, the sensitivity of B. anthracis to the PlyG lysin, or the selective amplification of environmental phages cocultured with B. anthracis. Findings from this work indicate that B. cereus sensu lato and its phages are common inhabitants of earthworm intestines.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacillus Phages / classification
  • Bacillus Phages / genetics
  • Bacillus Phages / growth & development
  • Bacillus Phages / isolation & purification*
  • Bacillus anthracis / classification
  • Bacillus anthracis / genetics
  • Bacillus anthracis / isolation & purification*
  • Bacillus anthracis / virology*
  • Bacillus cereus / virology
  • Cluster Analysis
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / genetics
  • DNA, Viral / chemistry
  • DNA, Viral / genetics
  • Gastrointestinal Tract / microbiology
  • Gastrointestinal Tract / virology
  • Metagenome
  • Molecular Sequence Data
  • Oligochaeta / microbiology*
  • Oligochaeta / virology*
  • Pennsylvania
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA
  • Viral Proteins / genetics

Substances

  • DNA, Bacterial
  • DNA, Ribosomal
  • DNA, Viral
  • RNA, Ribosomal, 16S
  • Viral Proteins

Associated data

  • GENBANK/GU142940
  • GENBANK/GU142941
  • GENBANK/GU142942
  • GENBANK/GU142943
  • GENBANK/GU142944
  • GENBANK/GU142945
  • GENBANK/GU142946
  • GENBANK/GU142947
  • GENBANK/GU142948