Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities
Abstract Background Cronobacter sakazakii is a foodborne pathogen that causes septicemia, meningitis, and necrotizing enterocolitis in neonates and infants. The current research details the full genome sequences of two extremely persistent C. sakazakii strains (H322 and GK1025B) isolated from powder...
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| Format: | Article |
| Language: | English |
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BMC
2022-06-01
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| Series: | Gut Pathogens |
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| Online Access: | https://doi.org/10.1186/s13099-022-00500-5 |
| _version_ | 1811258038458253312 |
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| author | Flavia J. Negrete Katie Ko Hyein Jang Maria Hoffmann Angelika Lehner Roger Stephan Séamus Fanning Ben D. Tall Gopal R. Gopinath |
| author_facet | Flavia J. Negrete Katie Ko Hyein Jang Maria Hoffmann Angelika Lehner Roger Stephan Séamus Fanning Ben D. Tall Gopal R. Gopinath |
| author_sort | Flavia J. Negrete |
| collection | DOAJ |
| description | Abstract Background Cronobacter sakazakii is a foodborne pathogen that causes septicemia, meningitis, and necrotizing enterocolitis in neonates and infants. The current research details the full genome sequences of two extremely persistent C. sakazakii strains (H322 and GK1025B) isolated from powdered infant formula (PIF) manufacturing settings. In addition, the genetic attributes associated with five plasmids, pH322_1, pH322_2, pGK1025B_1, pGK1025B_2, and pGK1025B_3 are described. Materials and Methods Using PacBio single-molecule real-time (SMRT®) sequencing technology, whole genome sequence (WGS) assemblies of C. sakazakii H322 [Sequence type (ST)83, clonal complex [CC] 83) and GK1025B (ST64, CC64) were generated. Plasmids, also sequenced, were aligned with phylogenetically related episomes to determine, and identify conserved and missing genomic regions. Results A truncated ~ 13 Kbp type 6 secretion system (T6SS) gene cluster harbored on virulence plasmids pH322_2 and pGK1025B_2, and a second large deletion (~ 6 Kbp) on pH322_2, which included genes for a tyrosine-type recombinase/integrase, a hypothetical protein, and a phospholipase D was identified. Within the T6SS of pH322_2 and pGK1025B_2, an arsenic resistance operon was identified which is in common with that of plasmids pSP291_1 and pESA3. In addition, PHASTER analysis identified an intact 96.9 Kbp Salmonella SSU5 prophage gene cluster in pH322_1 and pGK1025B_1 and showed that these two plasmids were phylogenetically related to C. sakazakii plasmids: pCS1, pCsa767a, pCsaC757b, pCsaC105731a. Plasmid pGK1025B_3 was identified as a novel conjugative Cronobacter plasmid. Furthermore, WGS analysis identified a ~ 16.4 Kbp type 4 secretion system gene cluster harbored on pGK1025B_3, which contained a phospholipase D gene, a key virulence factor in several host–pathogen diseases. Conclusion These data provide high resolution information on C. sakazakii genomes and emphasizes the need for furthering surveillance studies to link genotype to phenotype of strains from previous investigations. These results provide baseline data necessary for future in-depth investigations of C. sakazakii that colonize PIF manufacturing facility settings and genomic analyses of these two C. sakazakii strains and five associated plasmids will contribute to a better understanding of this pathogen's survival and persistence within various “built environments” like PIF manufacturing facilities. |
| first_indexed | 2024-04-12T18:07:07Z |
| format | Article |
| id | doaj.art-3562c82fd5124b3c889b1d543a948343 |
| institution | Directory Open Access Journal |
| issn | 1757-4749 |
| language | English |
| last_indexed | 2024-04-12T18:07:07Z |
| publishDate | 2022-06-01 |
| publisher | BMC |
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| series | Gut Pathogens |
| spelling | doaj.art-3562c82fd5124b3c889b1d543a9483432022-12-22T03:21:57ZengBMCGut Pathogens1757-47492022-06-0114111210.1186/s13099-022-00500-5Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilitiesFlavia J. Negrete0Katie Ko1Hyein Jang2Maria Hoffmann3Angelika Lehner4Roger Stephan5Séamus Fanning6Ben D. Tall7Gopal R. Gopinath8Center for Food Safety and Applied Nutrition, Office of Applied Research and Safety Assessment, U. S. Food and Drug AdministrationCenter for Food Safety and Applied Nutrition, Office of Applied Research and Safety Assessment, U. S. Food and Drug AdministrationCenter for Food Safety and Applied Nutrition, Office of Applied Research and Safety Assessment, U. S. Food and Drug AdministrationCenter for Food Safety and Applied Nutrition, Office of Regulatory Science, Food and Drug AdministrationInstitute for Food Safety and Hygiene, University of ZürichInstitute for Food Safety and Hygiene, University of ZürichWHO Collaborating Centre for Cronobacter, University College DublinCenter for Food Safety and Applied Nutrition, Office of Applied Research and Safety Assessment, U. S. Food and Drug AdministrationCenter for Food Safety and Applied Nutrition, Office of Applied Research and Safety Assessment, U. S. Food and Drug AdministrationAbstract Background Cronobacter sakazakii is a foodborne pathogen that causes septicemia, meningitis, and necrotizing enterocolitis in neonates and infants. The current research details the full genome sequences of two extremely persistent C. sakazakii strains (H322 and GK1025B) isolated from powdered infant formula (PIF) manufacturing settings. In addition, the genetic attributes associated with five plasmids, pH322_1, pH322_2, pGK1025B_1, pGK1025B_2, and pGK1025B_3 are described. Materials and Methods Using PacBio single-molecule real-time (SMRT®) sequencing technology, whole genome sequence (WGS) assemblies of C. sakazakii H322 [Sequence type (ST)83, clonal complex [CC] 83) and GK1025B (ST64, CC64) were generated. Plasmids, also sequenced, were aligned with phylogenetically related episomes to determine, and identify conserved and missing genomic regions. Results A truncated ~ 13 Kbp type 6 secretion system (T6SS) gene cluster harbored on virulence plasmids pH322_2 and pGK1025B_2, and a second large deletion (~ 6 Kbp) on pH322_2, which included genes for a tyrosine-type recombinase/integrase, a hypothetical protein, and a phospholipase D was identified. Within the T6SS of pH322_2 and pGK1025B_2, an arsenic resistance operon was identified which is in common with that of plasmids pSP291_1 and pESA3. In addition, PHASTER analysis identified an intact 96.9 Kbp Salmonella SSU5 prophage gene cluster in pH322_1 and pGK1025B_1 and showed that these two plasmids were phylogenetically related to C. sakazakii plasmids: pCS1, pCsa767a, pCsaC757b, pCsaC105731a. Plasmid pGK1025B_3 was identified as a novel conjugative Cronobacter plasmid. Furthermore, WGS analysis identified a ~ 16.4 Kbp type 4 secretion system gene cluster harbored on pGK1025B_3, which contained a phospholipase D gene, a key virulence factor in several host–pathogen diseases. Conclusion These data provide high resolution information on C. sakazakii genomes and emphasizes the need for furthering surveillance studies to link genotype to phenotype of strains from previous investigations. These results provide baseline data necessary for future in-depth investigations of C. sakazakii that colonize PIF manufacturing facility settings and genomic analyses of these two C. sakazakii strains and five associated plasmids will contribute to a better understanding of this pathogen's survival and persistence within various “built environments” like PIF manufacturing facilities.https://doi.org/10.1186/s13099-022-00500-5Cronobacter sakazakiiWhole genome sequencingPlasmidsBuilt environmentComplete genomesPHASTER |
| spellingShingle | Flavia J. Negrete Katie Ko Hyein Jang Maria Hoffmann Angelika Lehner Roger Stephan Séamus Fanning Ben D. Tall Gopal R. Gopinath Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities Gut Pathogens Cronobacter sakazakii Whole genome sequencing Plasmids Built environment Complete genomes PHASTER |
| title | Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities |
| title_full | Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities |
| title_fullStr | Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities |
| title_full_unstemmed | Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities |
| title_short | Complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent Cronobacter sakazakii strains ST83 H322 and ST64 GK1025B obtained from powdered infant formula manufacturing facilities |
| title_sort | complete genome sequences and genomic characterization of five plasmids harbored by environmentally persistent cronobacter sakazakii strains st83 h322 and st64 gk1025b obtained from powdered infant formula manufacturing facilities |
| topic | Cronobacter sakazakii Whole genome sequencing Plasmids Built environment Complete genomes PHASTER |
| url | https://doi.org/10.1186/s13099-022-00500-5 |
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