scholarly journals RNA Sequencing-Based Transcriptional Overview of Xerotolerance inCronobacter sakazakiiSP291

2018 ◽  
Vol 85 (3) ◽  
Author(s):  
Shabarinath Srikumar ◽  
Yu Cao ◽  
Qiongqiong Yan ◽  
Koenraad Van Hoorde ◽  
Scott Nguyen ◽  
...  
Keyword(s):  
Infant Formula ◽  
Mortality Rates ◽  
Secondary Response ◽  
Ecological Niches ◽  
Infant Food ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Rna Seq ◽  
Production Environment ◽  
Content Type

ABSTRACTCronobacter sakazakiiis a xerotolerant neonatal pathogen epidemiologically linked to powdered infant food formula, often resulting in high mortality rates. Here, we used transcriptome sequencing (RNA-seq) to provide transcriptional insights into the survival ofC. sakazakiiin desiccated conditions. Our RNA-seq data show that about 22% of the totalC. sakazakiigenes were significantly upregulated and 9% were downregulated during desiccation survival. When reverse transcription-quantitative PCR (qRT-PCR) was used to validate the RNA-seq data, we found that the primary desiccation response was gradually downregulated during the tested 4 hours of desiccation, while the secondary response remained constitutively upregulated. The 4-hour desiccation tolerance ofC. sakazakiiwas dependent on the immediate microenvironment surrounding the bacterial cell. The removal of Trypticase soy broth (TSB) salts and the introduction of sterile infant formula residues in the microenvironment enhanced the desiccation survival ofC. sakazakiiSP291. The trehalose biosynthetic pathway encoded byotsAandotsB, a prominent secondary bacterial desiccation response, was highly upregulated in desiccatedC. sakazakii.C. sakazakiiSP291 ΔotsABwas significantly inhibited compared with the isogenic wild type in an 8-hour desiccation survival assay, confirming the physiological importance of trehalose in desiccation survival. Overall, we provide a comprehensive RNA-seq-based transcriptional overview along with confirmation of the phenotypic importance of trehalose metabolism inCronobacter sakazakiiduring desiccation.IMPORTANCECronobacter sakazakiiis a pathogen of importance to neonatal health and is known to persist in dry food matrices, such as powdered infant formula (PIF) and its associated production environment. When infections are reported in neonates, mortality rates can be high. The success of this bacterium in surviving these low-moisture environments suggests thatCronobacterspecies can respond to a variety of environmental signals. Therefore, understanding those signals that aid the persistence of this pathogen in these ecological niches is an important step toward the development of strategies to reduce the risk of contamination of PIF. This research led to the identification of candidate genes that play a role in the persistence of this pathogen in desiccated conditions and, thereby, serve as a model target to design future strategies to mitigate PIF-associated survival ofC. sakazakii.

Alterations in the transcriptional landscape allows differential desiccation tolerance in different strains of Cronobacter sakazakii

2021 ◽  
Author(s):  
Yu Cao ◽  
Katherine Dever ◽  
Sathesh Kumar Sivasankaran ◽  
Scott V. Nguyen ◽  
Guerrino Macori ◽  
...  
Keyword(s):  
Infant Formula ◽  
Desiccation Tolerance ◽  
Growth Conditions ◽  
Evolutionary Strategies ◽  
Clinical Strain ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Rna Seq ◽  
Environmental Strain ◽  
Different Strains

Cronobacter sakazakii is a typical example of a xerotolerant bacterium. It is epidemiologically linked to low moisture foods like powdered infant formula (PIF) and is associated with high fatality rates among neonates. We characterized the xerotolerance in a clinically isolated strain, C. sakazakii ATCC™29544 T , and compared the desiccation tolerance with an environmental strain, C. sakazakii SP291, whose desiccation tolerance was previously characterized. We found that, although the clinical strain was desiccation-tolerant, the level of tolerance was compromised when compared to the environmental strain. RNA-seq based deep transcriptomic characterization identified a unique transcriptional profile in the clinical strain compared to what was already known for the environmental strain. As RNA-seq was also carried out in different TSB growth conditions, genes that were expressed specifically under desiccated conditions were identified and denoted as desiccation responsive genes (DRGs). Interestingly, these DRGs included transcriptomic factors like fnr , ramA, and genes associated with inositol metabolism, a phenotype as yet unreported in C. sakazakii . Further, the clinical strain did not express the proP gene, which was previously reported to be very important for desiccation survival and persistence. Interestingly, analysis of the plasmid genes showed that the iron metabolism in desiccated C. sakazakii ATCC™29544 T cells specifically involved the siderophore cronobactin encoded by the iucABCD genes. Confirmatory studies using qRT-PCR determined that, though the secondary desiccation response genes were upregulated in C. sakazakii ATCC™29544 T , the level of up-regulation was lower compared to that in C. sakazakii SP291. All these factors could collectively contribute to the compromised desiccation tolerance in the clinical strain. IMPORTANCE Cronobacter sakazakii has in past led to outbreaks, particularly associated with food that are low in moisture content. This species has adapted to survive in low water conditions and can survive in such environments for long periods. These characteristics have enabled the pathogen to contaminate powder infant formula, a food matrix with which the pathogen has been epidemiologically associated. Even though clinically adapted strains can also be isolated, there is no information on how the clinical strains adapt to low moisture environments. Our research assessed the adaptation of a clinically isolated strain to low moisture survival on sterile stainless steel coupons and compared the survival to a highly desiccation-tolerant environmental strain. We found that, even though the clinical strain is desiccation-tolerant, the rate of tolerance was compromised compared to the environmental strain. A deeper investigation using RNA-seq identified that the clinical strain used pathways different from that of the environmental strain to adapt to low moisture conditions. This shows that the adaptation to desiccation conditions, at least for C. sakazakii , is strain-specific and that different strains have used different evolutionary strategies for adaptation.

scholarly journals Draft Genome Sequence of Cronobacter sakazakii Cr268, Isolated from Infant Powder Formula

2019 ◽  
Vol 8 (46) ◽  
Author(s):  
Gal Zizelski Valenci ◽  
Mor Rubinstein ◽  
Reuven Afriat ◽  
Shira Rosencwaig ◽  
Zeev Dveyrin ◽  
...  
Keyword(s):  
Infant Formula ◽  
Necrotizing Enterocolitis ◽  
Genome Sequence ◽  
Premature Infants ◽  
Food Chain ◽  
Draft Genome ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Emerging Pathogen ◽  
Content Type

Cronobacter sakazakii is an emerging pathogen that causes meningitis, bacteremia, sepsis, and necrotizing enterocolitis in premature infants. Strain Cr268 was isolated from imported powdered infant formula in 2009 during routine microbial examination according to ISO-22964 (“Microbiology of the food chain—horizontal method for the detection of Cronobacter spp.”).

scholarly journals Draft Genome Sequences of Seven Cronobacter sakazakii Strains Carrying the mcr 9.1 Gene Isolated in Chile

2021 ◽  
Vol 10 (28) ◽  
Author(s):  
Julio Parra-Flores ◽  
Ondrej Holý ◽  
Francisca Riffo ◽  
Sarah Lepuschitz ◽  
Werner Ruppitsch ◽  
...  
Keyword(s):  
Infant Formula ◽  
Necrotizing Enterocolitis ◽  
Dairy Products ◽  
Draft Genome ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Genome Sequences ◽  
Content Type

Cronobacter sakazakii is a pathogen that causes severe diseases such as meningitis and necrotizing enterocolitis in infants under 12 months, associated with the consumption of contaminated rehydrated powdered infant formula (PIF). We present seven C. sakazakii genome sequences isolated from PIF and dairy products in Chile in 2017.

scholarly journals Draft Genome Sequences of Cronobacter muytjensii Cr150, Cronobacter turicensis Cr170, and Cronobacter sakazakii Cr611

2020 ◽  
Vol 9 (44) ◽  
Author(s):  
Gal Zizelski Valenci ◽  
Mor Rubinstein ◽  
Reuven Afriat ◽  
Shira Rosencwaig ◽  
Zeev Dveyrin ◽  
...  
Keyword(s):  
Infant Formula ◽  
Draft Genome ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Genome Sequences ◽  
Bioinformatics Analyses ◽  
New Members ◽  
Content Type

ABSTRACT The sequencing and bioinformatics analyses of isolates Cr150, Cr170, and Cr611 from powdered infant formula indicate that the three strains represent new members in the Cronobacter muytjensii, Cronobacter turicensis, and Cronobacter sakazakii groups, respectively.

scholarly journals Comparative Genotypic and Phenotypic Analysis of Cronobacter Species Cultured from Four Powdered Infant Formula Production Facilities: Indication of Pathoadaptation along the Food Chain

2015 ◽  
Vol 81 (13) ◽  
pp. 4388-4402 ◽  
Author(s):  
Qiongqiong Yan ◽  
Juan Wang ◽  
Jayanthi Gangiredla ◽  
Yu Cao ◽  
Marta Martins ◽  
...  
Keyword(s):  
Infant Formula ◽  
Sequence Type ◽  
Surveillance Program ◽  
Powdered Infant Formula ◽  
Clinical Environment ◽  
Production Environment ◽  
Dominant Type ◽  
Content Type ◽  
Production Environments ◽  
Production Facilities

ABSTRACTCronobacterspecies are opportunistic pathogens commonly found in the environment. Among the sevenCronobacterspecies,Cronobacter sakazakiisequence type 4 (ST-4) is predominantly associated with recorded cases of infantile meningitis. This study reports on a 26-month powdered infant formula (PIF) surveillance program in four production facilities located in distinct geographic regions. The objective was to identify the ST(s) in PIF production environments and to investigate the phenotypic features that support their survival. Of all 168Cronobacterisolates, 133 were recovered from a PIF production environment, 31 were of clinical origin, and 4 were laboratory type strains. Sequence type 1 (n= 84 isolates; 63.9%) was the dominant type in PIF production environments. The majority of these isolates clustered with an indistinguishable pulsotype and persisted for at least an 18-month period. Moreover, DNA microarray results identified two phylogenetic lineages among ST-4 strains tested. Thereafter, the ST-1 and -4 isolates were phenotypically compared. Differences were noted based on the phenotypes expressed by these isolates. The ST-1 PIF isolates produced stronger biofilms at both 28°C and 37°C, while the ST-4 clinical isolates exhibited greater swimming activity and increased binding to Congo red dye. Given the fact that PIF is a low-moisture environment and that the clinical environment provides for an interaction between the pathogen and its host, these differences may be consistent with a form of pathoadaptation. These findings help to extend our current understanding of the epidemiology and ecology ofCronobacterspecies in PIF production environments.

scholarly journals Genotyping and Source Tracking of Cronobacter sakazakii and C. malonaticus Isolates from Powdered Infant Formula and an Infant Formula Production Factory in China

2015 ◽  
Vol 81 (16) ◽  
pp. 5430-5439 ◽  
Author(s):  
Peng Fei ◽  
Chaoxin Man ◽  
Binbin Lou ◽  
Stephen J. Forsythe ◽  
Yunlei Chai ◽  
...  
Keyword(s):  
Infant Formula ◽  
Source Tracking ◽  
Neonatal Meningitis ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Sequence Analyses ◽  
Content Type ◽  
Fluidized Bed Drying ◽  
Contamination Routes ◽  
Sequence Types

ABSTRACTCronobacterspp. (formerly defined asEnterobacter sakazakii) are opportunistic bacterial pathogens of both infants and adults. In this study, we analyzed 70Cronobacterisolates from powdered infant formula (PIF) and an infant formula production facility in China to determine possible contamination routes. The strains were profiled by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), PCR-based O-antigen serotyping, andompAandrpoBsequence analyses. The isolates were primarilyCronobacter sakazakii(66/70) orCronobactermalonaticus(4/70). The strains were divided into 38 pulsotypes (PTs) using PFGE and 19 sequence types (STs) by MLST. In contrast,rpoBandompAsequence analyses divided the strains into 10 overlapping clusters each. PCR serotyping of the 66C. sakazakiiand 4C. malonaticusstrains resulted in the identification of fourC. sakazakiiserotypes (O1, O2, O4, and O7) and a singleC. malonaticusserotype, O2. The dominantC. sakazakiisequence types from PIF and an infant formula production factory in China wereC. sakazakiiclonal complex 4 (CC4) (n= 19), ST1 (n= 14), and ST64 (n= 11).C. sakazakiiCC4 is a clonal lineage strongly associated with neonatal meningitis. In the process of manufacturing PIF, the spray-drying, fluidized-bed-drying, and packing areas were the main areas withCronobactercontamination.C. sakazakiistrains with the same pulsotypes (PT3 and PT2) and sequence types (ST1 and ST64) were isolated both from processing equipment and from the PIF finished product.

Genetic Diversity of Cronobacter sakazakii Isolates Collected from a Swiss Infant Formula Production Facility

2013 ◽  
Vol 76 (5) ◽  
pp. 883-887 ◽  
Author(s):  
ANDREA MÜLLER ◽  
ROGER STEPHAN ◽  
CLAUDIA FRICKER-FEER ◽  
ANGELIKA LEHNER
Keyword(s):  
Infant Formula ◽  
Monitoring Program ◽  
Sequence Type ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Production Facility ◽  
Pfge Analysis ◽  
O Antigen ◽  
Selection For ◽  
Prevalent Species

In this study, 141 Cronobacter isolates that were collected based on a hygienic monitoring program performed in a powdered infant formula production facility in Switzerland between September 2011 and October 2012 were further characterized. Isolates were identified to the species level by molecular methods, and strains of Cronobacter sakazakii were further subtyped by applying PCR-based O-antigen serotyping, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). C. sakazakii was the most prevalent species identified (93.6%). Among this collection of isolates, representatives of all but one O-antigen serotype (serotype O5) were recognized. MLST analysis of 19 selected isolates revealed that most of the typeable isolates belonged to sequence type (ST) 4. Correlations between ST4 and serotype O2 and between ST83 and serotype O7 were observed. PFGE analysis revealed clusters with multiple isolates, including strains from samples collected at different time points and sampling sources. Generally, the observed heterogeneity among strains collected over the 13 months of the monitoring program was high, suggesting a constant flux among strains rather than a selection for persisting organisms.

scholarly journals Genome Sequences of Malonate-PositiveCronobacter sakazakiiSerogroup O:2, Sequence Type 64 Strains CDC 1121-73 and GK1025, Isolated from Human Bronchial Wash and a Powdered Infant Formula Manufacturing Plant

2016 ◽  
Vol 4 (6) ◽  
Author(s):  
H. R. Chase ◽  
G. R. Gopinath ◽  
J. Gangiredla ◽  
I. R. Patel ◽  
M. H. Kothary ◽  
...  
Keyword(s):  
Infant Formula ◽  
Draft Genome ◽  
Sequence Type ◽  
Powdered Infant Formula ◽  
Manufacturing Plant ◽  
Cronobacter Sakazakii ◽  
Genome Sequences ◽  
Manufacturing Facility ◽  
Bronchial Wash

We introduce draft genome sequences of strains CDC1121-73 (human bronchial wash isolate) and GK1025 (powdered infant formula manufacturing facility isolate), which are both malonate-positiveCronobacter sakazakiiserogroup O:2, sequence type 64. Assemblies for these strains have sizes of 4,442,307 and 4,599,266 bp and % G+C contents of 56.9 and 56.7, respectively.

scholarly journals Molecular characterization of Cronobacter sakazakii isolated from different herbal teas and mixtures in Serbia

Genetika ◽  
2017 ◽  
Vol 49 (3) ◽  
pp. 921-934
Author(s):  
Dragana Josic ◽  
Marija Stojanovic ◽  
Zorica Lepsanovic ◽  
Vera Katic
Keyword(s):  
Molecular Characterization ◽  
Infant Formula ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii ◽  
Safety Risks ◽  
High Prevalence ◽  
Sequences Analysis ◽  
Human Infections ◽  
Rrna Sequences

Cronobacter sakazakii is an important cause of human infections that can be serious and even fatal among premature neonates and immunocompromised adults or infants. Because of its high tolerance to osmotic stress, C. sakazakii is frequently isolated from dried foods, such as powdered infant formula and herbal teas. The aim of investigation was detection, identification and molecular characterization of Cronobacter sakazakii isolates from infant formula and various herbal teas collected from Serbian market and tested for import control. C. sakazakii was not detected in any of the 360 analysed samples of powdered infant formula. However, 192 out of 520 samples of herbal teas tested were positive for C. sakazakii (37.1%). The high prevalence was observed in teas for children (51.6%) and in ?baby? teas (44.1%), followed by medicinal teas (38%). The largest one-herb-teas group (221 samples) contained 72 C. sakazakii-positive samples (32.6%) and involved Sennae folium, Althaeae radix, Menthae piperitae folium, Chamomilae flos and Urticae folium teas. Molecular characterization of isolated C. sakazakii from different herbal teas by rep-PCR, RAPD and 16S rRNA sequences analysis showed the high similarity to C. sakazakii NCTC 8155. Knowing this strain as one of the most pathogenic clinical strains, our results raise concern about the safety risks these foods pose to immunocompromised and healthy consumers, especially for babies and children.

Heat resistance of Cronobacter sakazakii DPC 6529 and its behavior in reconstituted powdered infant formula

2015 ◽  
Vol 69 ◽  
pp. 401-409 ◽  
Author(s):  
Juan-Pablo Huertas ◽  
Avelino Álvarez-Ordóñez ◽  
Ruth Morrissey ◽  
Maria Ros-Chumillas ◽  
Maria-Dolores Esteban ◽  
...  
Keyword(s):  
Heat Resistance ◽  
Infant Formula ◽  
Powdered Infant Formula ◽  
Cronobacter Sakazakii
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