TY - JOUR
T1 - Characterization of Cronobacter sakazakii Strains Originating from Plant-Origin Foods Using Comparative Genomic Analyses and Zebrafish Infectivity Studies
AU - Jang, Hyein
AU - Eshwar, Athmanya
AU - Lehner, Angelika
AU - Gangiredla, Jayanthi
AU - Patel, Isha R.
AU - Beaubrun, Junia Jean Gilles
AU - Chase, Hannah R.
AU - Negrete, Flavia
AU - Finkelstein, Samantha
AU - Weinstein, Leah M.
AU - Ko, Katie
AU - Addy, Nicole
AU - Ewing, Laura
AU - Woo, Jungha
AU - Lee, Youyoung
AU - Seo, Kunho
AU - Jaradat, Ziad
AU - Srikumar, Shabarinath
AU - Fanning, Séamus
AU - Stephan, Roger
AU - Tall, Ben D.
AU - Gopinath, Gopal R.
N1 - Funding Information:
We thank the student internship programs sponsored by the Offices of International Affairs of Gachon University, Seongnam-si, Republic of Korea for supporting student interns: J.W. and Y.L. We thank the University of Maryland, Joint Institute for Food Safety and Applied Nutrition (JIFSAN) for supporting JIFSAN interns S.F., F.N., K.K. and L.M.W. through a cooperative agreement with the FDA, #FDU001418. We also thank the Oak Ridge Institute for Science and Education of Oak Ridge (ORISE), Tennessee for sponsoring research fellows H.R.C., N.A. and H.J. All authors read and approved the Acknowledgements.
Publisher Copyright:
© 2022 by the authors.
PY - 2022/7
Y1 - 2022/7
N2 - Cronobacter sakazakii continues to be isolated from ready-to-eat fresh and frozen produce, flours, dairy powders, cereals, nuts, and spices, in addition to the conventional sources of powdered infant formulae (PIF) and PIF production environments. To understand the sequence diversity, phylogenetic relationship, and virulence of C. sakazakii originating from plant-origin foods, comparative molecular and genomic analyses, and zebrafish infection (ZI) studies were applied to 88 strains. Whole genome sequences of the strains were generated for detailed bioinformatic analysis. PCR analysis showed that all strains possessed a pESA3-like virulence plasmid similar to reference C. sakazakii clinical strain BAA-894. Core genome analysis confirmed a shared genomic backbone with other C. sakazakii strains from food, clinical and environmental strains. Emerging nucleotide diversity in these plant-origin strains was highlighted using single nucleotide polymorphic alleles in 2000 core genes. DNA hybridization analyses using a pan-genomic microarray showed that these strains clustered according to sequence types (STs) identified by multi-locus sequence typing (MLST). PHASTER analysis identified 185 intact prophage gene clusters encompassing 22 different prophages, including three intact Cronobacter prophages: ENT47670, ENT39118, and phiES15. AMRFinderPlus analysis identified the CSA family class C β-lactamase gene in all strains and a plasmid-borne mcr-9.1 gene was identified in three strains. ZI studies showed that some plant-origin C. sakazakii display virulence comparable to clinical strains. Finding virulent plant-origin C. sakazakii possessing significant genomic features of clinically relevant STs suggests that these foods can serve as potential transmission vehicles and supports widening the scope of continued surveillance for this important foodborne pathogen.
AB - Cronobacter sakazakii continues to be isolated from ready-to-eat fresh and frozen produce, flours, dairy powders, cereals, nuts, and spices, in addition to the conventional sources of powdered infant formulae (PIF) and PIF production environments. To understand the sequence diversity, phylogenetic relationship, and virulence of C. sakazakii originating from plant-origin foods, comparative molecular and genomic analyses, and zebrafish infection (ZI) studies were applied to 88 strains. Whole genome sequences of the strains were generated for detailed bioinformatic analysis. PCR analysis showed that all strains possessed a pESA3-like virulence plasmid similar to reference C. sakazakii clinical strain BAA-894. Core genome analysis confirmed a shared genomic backbone with other C. sakazakii strains from food, clinical and environmental strains. Emerging nucleotide diversity in these plant-origin strains was highlighted using single nucleotide polymorphic alleles in 2000 core genes. DNA hybridization analyses using a pan-genomic microarray showed that these strains clustered according to sequence types (STs) identified by multi-locus sequence typing (MLST). PHASTER analysis identified 185 intact prophage gene clusters encompassing 22 different prophages, including three intact Cronobacter prophages: ENT47670, ENT39118, and phiES15. AMRFinderPlus analysis identified the CSA family class C β-lactamase gene in all strains and a plasmid-borne mcr-9.1 gene was identified in three strains. ZI studies showed that some plant-origin C. sakazakii display virulence comparable to clinical strains. Finding virulent plant-origin C. sakazakii possessing significant genomic features of clinically relevant STs suggests that these foods can serve as potential transmission vehicles and supports widening the scope of continued surveillance for this important foodborne pathogen.
KW - Cronobacter sakazakii
KW - draft genomes
KW - dried foods
KW - plant-origin foods
KW - whole genome sequencing (WGS)
UR - http://www.scopus.com/inward/record.url?scp=85136357703&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85136357703&partnerID=8YFLogxK
U2 - 10.3390/microorganisms10071396
DO - 10.3390/microorganisms10071396
M3 - Article
AN - SCOPUS:85136357703
SN - 2076-2607
VL - 10
JO - Microorganisms
JF - Microorganisms
IS - 7
M1 - 1396
ER -