A simple and sensitive aptasensor with rolling circle amplification for viable Cronobacter sakazakii detection in powdered infant formula

Author(s):  
Ju Liu ◽  
Guoyang Xie ◽  
Qin Xiong ◽  
Dan Mu ◽  
Hengyi Xu
2018 ◽  
Vol 85 (3) ◽  
Author(s):  
Shabarinath Srikumar ◽  
Yu Cao ◽  
Qiongqiong Yan ◽  
Koenraad Van Hoorde ◽  
Scott Nguyen ◽  
...  

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.


2013 ◽  
Vol 76 (5) ◽  
pp. 883-887 ◽  
Author(s):  
ANDREA MÜLLER ◽  
ROGER STEPHAN ◽  
CLAUDIA FRICKER-FEER ◽  
ANGELIKA LEHNER

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.


2016 ◽  
Vol 4 (6) ◽  
Author(s):  
H. R. Chase ◽  
G. R. Gopinath ◽  
J. Gangiredla ◽  
I. R. Patel ◽  
M. H. Kothary ◽  
...  

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.


Genetika ◽  
2017 ◽  
Vol 49 (3) ◽  
pp. 921-934
Author(s):  
Dragana Josic ◽  
Marija Stojanovic ◽  
Zorica Lepsanovic ◽  
Vera Katic

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.


2015 ◽  
Vol 69 ◽  
pp. 401-409 ◽  
Author(s):  
Juan-Pablo Huertas ◽  
Avelino Álvarez-Ordóñez ◽  
Ruth Morrissey ◽  
Maria Ros-Chumillas ◽  
Maria-Dolores Esteban ◽  
...  

2015 ◽  
Vol 78 (9) ◽  
pp. 1651-1655 ◽  
Author(s):  
DONG-HYEON KIM ◽  
JUNG-WHAN CHON ◽  
IL-BYEONG KANG ◽  
HYUNSOOK KIM ◽  
HONG-SEOK KIM ◽  
...  

Kefir is a type of fermented milk containing lactic and acetic acid bacteria and yeast. In this study, we evaluated the antimicrobial activity of kefir supernatant against Cronobacter sakazakii in powdered infant formula (PIF). In a spot-on-lawn test, the growth of 20 C. sakazakii strains, including 10 clinical and 10 food isolates, was completely inhibited in the presence of kefir supernatant. Significant differences in the diameters of inhibition zones were observed upon treatment with kefir compared with the results for Lactobacillus kefiri and Candida kefyr culture supernatants or solutions of lactic and acetic acid and ethyl alcohol in the agar well diffusion test (P < 0.05). The addition of 100 μl of kefir supernatant to 1 ml of nutrient broth completely inhibited the growth of C. sakazakii, as evaluated by spectrophotometry. The antimicrobial activity of kefir supernatant in experimentally contaminated PIF was also tested; we found no viable C. sakazakii cells remaining in PIF rehydrated with 30% kefir supernatant solution for 1 h, demonstrating that the antimicrobial activity of kefir supernatant against C. sakazakii could be applied in real food samples.


Author(s):  
Yu Cao ◽  
Katherine Dever ◽  
Sathesh Kumar Sivasankaran ◽  
Scott V. Nguyen ◽  
Guerrino Macori ◽  
...  

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.


2019 ◽  
Vol 7 (3) ◽  
pp. 77 ◽  
Author(s):  
Monica Henry ◽  
Aliyar Fouladkhah

Previously known as Enterobacter sakazakii from 1980 to 2007, Cronobacter sakazakii is an opportunistic bacterium that survives and persists in dry and low-moisture environments, such as powdered infant formula. Although C. sakazakii causes disease in all age groups, infections caused by this pathogen are particularly fatal in infants born premature and those younger than two months. The pathogen has been isolated from various environments such as powdered infant formula manufacturing facilities, healthcare settings, and domestic environments, increasing the chance of infection through cross-contamination. The current study discusses the outbreak history of C. sakazakii and the ability of the microorganism to produce biofilms on biotic and abiotic surfaces. The study further discusses the fate of the pathogen in low-moisture environments, articulates preventive measures for healthcare providers and nursing parents, and delineates interventions that could be utilized in infant formula manufacturing to minimize the risk of contamination with Cronobacter sakazakii.


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