2528 | wileyonlinelibrary.com/journal/jam J Appl Microbiol. 2022;133:2528–2546. © 2022 Society for Applied Microbiology. INTRODUCTION The genus Cronobacter comprises seven spe- cies: Cronobacter sakazakii, Cronobacter malonati- cus, Cronobacter muytjensii, Cronobacter turicensis, Cronobacter dublinensis, Cronobacter universalis and Cronobacter condimenti (Iversen et al., 2007, 2008; Joseph et al., 2011; Stephan et al., 2014). Of the seven species, C. sakazakii is the primary pathogen causing most illnesses in infants as well as in elderly individuals (Jang, Woo, et al., 2018). Unexpectedly, surveillance data obtained by analysing hospital records through the USA FoodNet sur- veillance system reported that there is a higher percent- age of Cronobacter infections observed in adults than in Received: 14 April 2022 | Revised: 14 July 2022 | Accepted: 15 July 2022 DOI: 10.1111/jam.15723 ORIGINAL ARTICLE Virulence, antimicrobial susceptibility and phylogenetic analysis of Cronobacter sakazakii isolates of food origins from Jordan Ziad W. Jaradat 1 | Waseem A. Al-Mousa 1 | Ahmed M. Elbetieha 1 | Qutaiba O. Ababneh 1 | Anas A. Al-Nabulsi 2 | Hyein Jang 3 | Jayanthi Gangiredla 3 | Isha R. Patel 3 | Gopal R. Gopinath 3 | Ben D. Tall 3 1 Department of Biotechnology and Genetic Engineering, Jordan University of Science and Technology, Irbid, Jordan 2 Department of Nutrition and Food Technology, Jordan University of Science and Technology, Irbid, Jordan 3 US Food and Drug Administration, Center for Food Safety and Applied Nutrition, Laurel, Maryland, USA Correspondence Ziad W. Jaradat, Department of Biotechnology and Genetic Engineering, Jordan University of Science and Technology, Irbid 22110, P. O Box 3030, Jordan. Email: jaradatz@just.edu.jo Funding information Deanship of Research, Jordan University of Science and Technology, Grant/Award Number: 20100054; US FDA; Jordan University of Science and Technology Abstract Aims: The aim was to characterize a collection of Cronobacter sakazakii isolates collected from various origins in Jordan. Methods and Results: The isolates were characterized using 16S rRNA sequencing, DNA microarray, multi-locus sequence typing (MLST), O-serotyping, virulence gene identification and antibiotic susceptibility testing. The identities and phylogenetic relatedness revealed that C. sakazakii sequence type 4 (ST4) and Csak O:1 serotype were the most prevalent STs and serovars amongst these C. sakazakii strains. PCR screening of putative virulence genes showed that the siderophore-interacting protein gene (sip) and iron acquisition gene clusters (eitCBAD and iucABCD/iutA) were the most detected genes with noticeable variability in the type 6 secretion system (T6SS) and filamentous hemagglutinin/adhesion (FHA) gene loci. The antibiotic resistance profiles revealed that the majority of the isolates were susceptible to all antibiotics used despite harbouring a class C β-lactamase resistance gene. Conclusions: The results described in this report provide additional insights about the considerable genotypic and phenotypic heterogeneity within C. sakazakii. Significance and Impact of the Study: The information reported in this study might be of great value in understanding the origins of C. sakazakii isolates, in addition to their diversity and variability, which might be helpful in preventing future outbreaks of this pathogen. KEYWORDS antibiotic resistance, C. sakazakii, phylogenetic analysis, virulence genes Downloaded from https://academic.oup.com/jambio/article/133/4/2528/6989083 by guest on 19 January 2023