Skip to main content
Genome Announcements logoLink to Genome Announcements
. 2015 Dec 17;3(6):e01480-15. doi: 10.1128/genomeA.01480-15

Next-Generation Whole-Genome Sequencing of Eight Strains of Bacillus cereus, Isolated from Food

Antonina O Krawczyk a,c, Anne de Jong a,c, Robyn T Eijlander a,c, Erwin M Berendsen a,b,c, Siger Holsappel a, Marjon H J Wells-Bennik b,c, Oscar P Kuipers a,c,
PMCID: PMC4683234  PMID: 26679589

Abstract

Bacillus cereus can contaminate food and cause emetic and diarrheal foodborne illness. Here, we report whole-genome sequences of eight strains of B. cereus, isolated from different food sources.

GENOME ANNOUNCEMENT

Bacillus cereus is a mesophilic or psychrotrophic, spore-forming bacterium commonly present in soil (1). It occurs in the rhizosphere of plants (2, 3) and as a part of animal intestinal microflora (4). It is opportunistically pathogenic and leads to various infections, including: local infections of wounds, or an eye; bacteremia and septicemia; respiratory infections; central nervous system infections; pericarditis; and endocarditis (5). Due to its presence in soil and production of spores, B. cereus often contaminates various food products. Consumption of foods with high levels of B. cereus may result in two types of foodborne illness: emetic or diarrheal. The emetic type, characterized by vomiting and nausea, is induced by the cereulide toxin produced by cells growing in food (68). The diarrheal illness is caused by enterotoxins, including hemolysin BL, cytotoxin K, and nonhemolytic enterotoxin, which are produced by B. cereus cells in the small intestine (6, 7). B. cereus is closely related to Bacillus anthracis, the causative agent of anthrax, and to the insect pathogen, Bacillus thuringiensis (9).

Eight strains of B. cereus, isolated from different food sources were sequenced by next-generation whole-genome sequencing. The strains were grown at 30°C with shaking at 220 rpm in heart infusion (BHI) broth (Difco). The overnight cultures were diluted in fresh medium to the optical density at 600 nm (OD600) and harvested by centrifugation at 5,000 relative centrifugal force (RCF). Subsequently, total DNA was isolated by phenol-chloroform extraction as described previously (10). The isolated DNA was sheared to 500-bp fragments in the Covaris (KBioscience) ultrasone device for preparing the NGS library preps using the paired-end NEB NExtGen library preparation kit. The libraries were 101-base paired-end sequenced on an Illumina HiSeq2000. Subsequently, Velvet (11) was used to perform a de novo paired-end assembly on each genome resulting in the draft genome sequences. The RAST server (12) and BAGEL3 (13) were used to annotate the genomes and to identify putative bacteriocin gene clusters, respectively.

Nucleotide sequence accession numbers.

The genome sequence of the eight Bacillus cereus strains have been deposited as whole-genome shotgun projects at DDBJ/EMBL/GenBank under the accession numbers listed in Table 1.

TABLE 1.

Sequenced strains and their sourcesa

B. cereus strain Source Accession no.
B4077 Chilled dessert LCYI00000000
B4078 Food, undefined LCYJ00000000
B4080 Dried onion LCYK00000000
B4086 Boiled rice LCYL00000000
B4087 Pea soup LCYM00000000
B4147 Cereals, pasta and pastries LCYN00000000
B4153 Dairy products LCYO00000000
B4158 Vegetables LCYP00000000
a

B-numbers refer to the strain collection at NIZO food research and the University of Groningen (Molecular Genetics).

ACKNOWLEDGMENTS

We thank the NGS sequence facility of the University Medical Center of Groningen (UMCG) for performing the sequencing of the strains. We thank Top Institute for Food and Nutrition for contributing to the funding of the project in theme 3: Safety and Preservation.

Footnotes

Citation Krawczyk AO, de Jong A, Eijlander RT, Berendsen EM, Holsappel S, Wells-Bennik MHJ, Kuipers OP. 2015. Next-generation whole-genome sequencing of eight strains of Bacillus cereus, isolated from food. Genome Announc 3(6):e01480-15. doi:10.1128/genomeA.01480-15.

REFERENCES

  • 1.Ceuppens S, Boon N, Uyttendaele M. 2013. Diversity of Bacillus cereus group strains is reflected in their broad range of pathogenicity and diverse ecological lifestyles. FEMS Microbiol Ecol 84:433–450. doi: 10.1111/1574-6941.12110. [DOI] [PubMed] [Google Scholar]
  • 2.Halverson LJ, Clayton MK, Handelsman J. 1993. Population biology of Bacillus cereus UW85 in the rhizosphere of field-grown soybeans. Soil Biol Biochem 25:485–493. doi: 10.1016/0038-0717(93)90074-L. [DOI] [Google Scholar]
  • 3.Zhang S, Liao S, Yu X, Lu H, Xian J, Guo H, Wang A, Xie J. 2015. Microbial diversity of mangrove sediment in Shenzhen Bay and gene cloning, characterization of an isolated phytase-producing strain of SPC09 B. cereus. Appl Microbiol Biotechnol 99:5339–5350. doi: 10.1007/s00253-015-6405-8. [DOI] [PubMed] [Google Scholar]
  • 4.Margulis L, Jorgensen JZ, Dolan S, Kolchinsky R, Rainey FA, Lo S-C. 1998. The Arthromitus stage of Bacillus cereus: intestinal symbionts of animals. Proc Natl Acad Sci USA 95:1236–1241. doi: 10.1073/pnas.95.3.1236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Drobniewski FA. 1993. Bacillus cereus and related species. Clin Microbiol Rev 6:324–338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Logan NA. 2012. Bacillus and relatives in foodborne illness. J Appl Microbiol 112:417–429. doi: 10.1111/j.1365-2672.2011.05204.x. [DOI] [PubMed] [Google Scholar]
  • 7.Stenfors Arnesen LP, Fagerlund A, Granum PE. 2008. From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev 32:579–606. doi: 10.1111/j.1574-6976.2008.00112.x. [DOI] [PubMed] [Google Scholar]
  • 8.Granum PE, Lund T. 2006. Bacillus cereus and its food poisoning toxins. FEMS Microbiol Lett 157:223–228. doi: 10.1111/j.1574-6968.1997.tb12776.x. [DOI] [PubMed] [Google Scholar]
  • 9.Rasko DA, Altherr MR, Han CS, Ravel J. 2005. Genomics of the Bacillus cereus group of organisms. FEMS Microbiol Rev 29:303–329. doi: 10.1016/j.fmrre.2004.12.005. [DOI] [PubMed] [Google Scholar]
  • 10.Krawczyk AO, Berendsen EM, Eijlander RT, de Jong A, Wells-Bennik MHJ, Kuipers OP. 2015. Draft genome sequences of four Bacillus thermoamylovorans strains isolated from milk and acacia gum, a food ingredient. Genome Announc 3(2):e00165-15. doi: 10.1128/genomeA.00165-15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Zerbino DR, Birney E. 2008. Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 18:821–829. doi: 10.1101/gr.074492.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Aziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, Formsma K, Gerdes S, Glass EM, Kubal M, Meyer F, Olsen GJ, Olson R, Osterman AL, Overbeek RA, McNeil LK, Paarmann D, Paczian T, Parrello B, Pusch GD, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O. 2008. The RAST Server: rapid annotations using subsystems technology. BMC Genomics 9:75. doi: 10.1186/1471-2164-9-75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.van Heel AJ, de Jong A, Montalbán-López M, Kok J, Kuipers OP. 2013. BAGEL3: automated identification of genes encoding bacteriocins and (non-) bactericidal posttranslationally modified peptides. Nucleic Acids Res 41:W448–W453. doi: 10.1093/nar/gkt391. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genome Announcements are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES