• The Korean Journal of Food And Nutrition
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• v.28 no.4
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• pp.728-736
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• 2015

Cronobacter species (Cronobacter spp.), previously known as Enterobacter sakazakii, are gram negative food borne pathogenic bacteria. They pose a very high risk of infection to neonates and immuno-compromised individuals and can affect the human central nervous system. Consequently, survivors often suffer from severe neurological impairment including hydrocephalus, quadriplegia, and developmental delays. Cronobacter spp. were not only isolated from plant food and products such as cereals, fruits, vegetables, legume products, herbs, and spices but also from animal source foods such as milk, meat, fish, and products made from these foods. Therefore, rapid detection of Cronobacter spp. is essential for food safety. Many detection methods have been developed since the Cronobacter spp. were first reported. However, the development of more rapid, sensitive, and easy-to-use detection methods for the Cronobacter spp. is required. In this review, our aim was to study and compare the available detection methods for Cronobacter spp., including culture-based, molecular biology-based, and immunology-based methods. This study will contribute to the development of new and rapid detection method for Cronobacter spp.

• Korean Journal of Veterinary Service
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• v.36 no.2
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• pp.67-71
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• 2013

Neonatal infection caused by Cronobacter species can result in serious illnesses such as bacteremia, septicemia, meningitis, and death in at-risk infants who are orally fed contaminated reconstituted powdered infant formulas. The objective of this study was to compare the virulence among three Cronobacter species strains by using an animal model for human neonatal Cronobacter species infections. We acquired timed-pregnant ICR mice and all owed them to give birth naturally. On postnatal day 3, each pup was administered orally a total dose of $1{\times}10^7$ CFU Cronobacter species strain 3439, CDC 1123-79, and 3231. Mice were observed twice daily for morbidity and mortality. At postnatal day 10, the remaining pups were euthanized, and brain, liver, and cecum were excised and analyzed for the presence of Cronobacter species. Cronobacter species were isolated from cecum and other tissues in inoculated mice. In the tissues of Cronobacter species infected mice, meningitis and gliosis were detected in the brain. In this study, we identified the virulence among Cronobacter species strains by using a neonatal mice model which was a very effective animal model for human neonatal Cronobacter species infections.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.5
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• pp.764-768
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• 2010

Three different isolation media for Cronobacter sakazakii have been recommended by Korea Food and Drug Administration from 2007. Performance comparison test was carried out between recommended Cronobacter sakazakii isolation medium. Chromogenic Enterobacter sakazakii agar (CESA) and Enterobacter sakazakii agar (ESA) produce more distinctive colonies having characteristic colors and appearance than Violet red bile glucose agar (VRBGA). The sensitivity and specificity of 3 different isolation media was checked. All 3 tested media showed 100% sensitivity when tested with 30 different Cronobacter sakazakii. The CESA and ESA showed 100% specificity when tested with Enterobacteriaceae except Cronobacter sakazakii, However, VRBGA did not show any specificity, showing inadequate selectivity compared to applicable Cronobacter sakazakii isolation medium. Artificially inoculated Cronobacter sakazakii to milk powder was easily recovered with 3 different isolation media and they all showed almost the same recovery activity.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.229-239
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• 2012

Cronobacter spp. (formerly Enterobacter sakazakii), a Gram-negative bacillus, is a rare cause of meningitis and central nervous system infections. In England, the first case infected by this organism occurred in 1958. By July 2008, approximately 120 documented cases of Cronobacter spp. infection and at least 27 deaths have been identified from all around the world in the published literature and in reports submitted by public health sectors. In 2007, it was proposed by European organizations that the original taxonomy of E. sakazakii would be revised, to consist of five new species moved to a new genus, and identified as "Cronobacter". E. sakazakii has thus now been reclassified as 6 separate species in the new genus, Cronobacter, gen. nov., within the Enterobacteriaceae family. The new species are presently Cronobacter sakazakii, C. turicensis, C. malonaticus, C. muytjensii, and C. dublinensis; the sixth species is identified simply as genomospecies I, as currently including only two representative strains. The objectives of this review are to provide insight on (1) the classification and taxonomy of Cronobacter spp., (2) its clinical etiology and pathogenicity, (3) prequency of Cronobacter spp. in different categories of ready-to-eat food other than infant formula, (4) methods for detecting, isolating and typing Cronobacter spp., and (5) recent research trends for detecting Cronobacter spp.

• Food Science of Animal Resources
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• v.38 no.2
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• pp.376-390
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• 2018

Cronobacter species have been associated with disease outbreaks and sporadic infections, particularly in premature and immunocompromised infants. Cronobacter species can cause foodborne infections such as neonatal meningitis, septicaemia and necrotising enterocolitis. Accordingly, there is an urgent need to control and monitor the Cronobacter species in food, especially in powdered infant formula (PIF) and other baby foods. Therefore, in this review, the isolation and prevalence of Cronobacter species in infant food including PIF and the recent advance of detection methods are discussed for the better understanding on the current research status of Cronobacter species.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.757-762
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• 2010

Cronobacter is a major foodborne pathogen in powdered infant formula and can lead to serious developmental after-effect and death to infants. The contamination of Cronobacter may be a high risk for the powdered foods. To isolate and identify Cronobacter from the powdered foods such as powdered infant formula and Saengsik in Korea, a conventional culture method, rapid identification system, PCR, and 16S rDNA sequencing were performed. As the results of isolation, seven Cronobacter spp. were isolated from seven out of 102 powdered infant formulas and 41 Cronobacter were isolated from 41 out of 86 Saengsiks. Forty-eight Cronobacter isolates were identified to be C. sakazakii and C. dublenisis by 16S rDNA sequence analysis. Most of the isolates were C. sakazakii and 13% of the isolates were C. dublinesis. One fourth of the C. sakazakii isolates showed different biochemical characteristics of negative nitrate reduction and nonmotility activities compared with the other strains reported previously.

• Journal of Dairy Science and Biotechnology
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• v.39 no.3
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• pp.113-120
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• 2021

Although the number of incidences of illness caused by ingestion of the bacterial pathogen Enterobacter sakazakii (Cronobacter spp.) has dramatically declined, there remains a need for a robust isolation method to recover this microbe from powdered infant formula (PIF). The current method described in the FDA's Bacteriological Analytical Manual requires multiple steps, and 3-4+ days for complete analysis of PIF isolated E. sakazakii (Cronobacter spp.). We describe a bacteriological method including a one-step enrichment followed by plating on chromogenic agar for presumptive identification of E. sakazakii (Cronobacter spp.). Suspected colonies are confirmed by either biochemical analyses, or a Real-Time PCR-based assay. Using this method, E. sakazakii (Cronobacter spp.) in PIF can be isolated and identified within one day (24 hours).

• Food Science of Animal Resources
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• v.30 no.3
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• pp.479-486
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• 2010

Tolerance against heat, sanitizers, and antibiotics of 112 Cronobacter isolates classified by desiccation was determined to permit effective biocontrol in powdered foods. The isolates were classified into three groups: dry-tolerant (n=37), dry-sensitive (n=7), and dry-intermediate (n=68). The strains that were highly tolerant to drying also showed high heat tolerance that they seemed to have high tolerance to heat after dry stress in powdered foods like infant formula. Sodium hypochlorite and benzalkonium chloride concentrations necessary to achieve a 5-log reduction in viable counts (CFU/mL) were 15-25 ppm and 5-15 ppm, respectively. However, there was little difference of the efficacy of these sanitizers between dry-sensitive and -tolerant strains for planktonic cells suspended in 3% albumin. The minimal inhibition concentration (MIC) of $\beta$-lactam ampicillin was 64-128 ppm for 90% of the strains. The isolates were consistently sensitive to kanamycin and naldixic acid (MIC=4 ppm). Dry-tolerant strains displayed more antibiotic resistance than dry-sensitive strains. The results indicate that dry-tolerant Cronobacter isolates often possess heat and antibiotic resistance, indicated the need for potent sterilization treatments of powdered foods.

• Korean Journal of Food Science and Technology
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• v.41 no.6
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• pp.681-686
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• 2009

Cronobacter spp. (Enterobacter sakazakii) is known to be highly resistant to dry conditions than any other Enterobacteriaeae. In this study, one hundred and ten Korean Cronobacter isolates were characterized to find out their survival characteristics under conditions of desiccation and humidity. Thirty percentage strains of the isolates showed high resistance to desiccation exposed on the metal surface for eight hours by half survival of the initial number, whileas less than 10% strains showed dry sensitivity by less one log scale survival among seven log scales. Finally, more than 90% of the strains consisted of dry-resistant and dry-intermediate groups. The same tendencies were evident in a 15-day exposure. Dry-resistant and intermediate strain groups showed 3 log scale survival among 5 log initial numbers in the powdered infant formula for 30 days, which were more resistant than on the above metal surface exposed. So, almost the isolate strains showed high resistance to dry condition. Dry-resistant and intermediate groups exposed on the metal surface formed a biofilm at the beginning, and the dry-sensitive group showed biofilm formation mainly only after a 7-day exposure. However, without a time difference in formation of biofilm, the dry-resistant and sensitive isolates seemed to similar biofilm formation activity. Most of the isolates showed very low survival at 75% relative humidity in 48 hours; however, they showed high resistance by 60% survival at 40% relative humidity. The Cronobacter isolates showed high resistance to desiccation on the metal surface and in the powdered infant formula, but low survival at high relative humidity. Therefore, high humidity may be a control method for Cronobacter in food processing environments.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.702-708
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• 2009

We investigated biofilm formation in various media, growth in low pH, and the hemolytic activity of 14 strains of Cronobacter spp. (Enterobacter sakazakii) isolated from a variety of foods including powdered infant formula (n=75), infant cereal (n=100), honey (n=30), and other infant foods (n=100) in Korea. The Cronobacter spp. adhered and formed biofilms on polyethylene, and a greater extent of biofilm was observed in nutrient-rich media. No clear difference in biofilm-forming ability was noted among the media constituents and the pattern of biofilm formation was strain-dependent. Seven strains out of 14 strains (50%) grew at pH 4.1, indicating that the acid resistance of these Cronobacter spp. isolated in Korea was relatively low. Hemolytic activity was not observed in any of the strains. This study provides basic information for the physiological and biochemical characteristics of Cronobacter spp. isolated from a variety of infant foods in Korea.