• 제목/요약/키워드: Enterobacter sakazakii (Cronobacter spp.)

검색결과 13건 처리시간 0.02초

Rates of Recovery of Enterobacter sakazakii (Cronobacter spp.) from Powdered Infant Formula Using Both a Chromogenic Agar and Real-Time PCR : A Preliminary Study

  • Song, Kwang-Young;Seo, Kun-Ho;Chon, Jung-Whan
    • Journal of Dairy Science and Biotechnology
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    • 제39권3호
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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).

Evaluation of Commercial Disinfectants for Efficacy at Inactivating Enterobacter sakazakii (Cronobacter spp.) in Water: A Preliminary Study

  • Chon, Jung-Whan;Seo, Kun-Ho;Kim, Binn;Her, Jekang;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • 제39권3호
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    • pp.104-112
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    • 2021
  • This study was conducted to evaluate the efficacy of commercial disinfectants at inactivating Enterobacter sakazakii (Cronobacter spp.) in water. Disinfectant I contained 6.15% sodium hypochlorite, and disinfectant II contained both 2.25% n-alkyl dimethylbenzyl ammonium chloride and 2.25% n-alkyl ethylbenzyl ammonium chloride. Disinfectant I was added to distilled water to obtain a range of residual chloride concentrations at 50 ppm intervals with a maximum of 1-1,000 ppm. Disinfectant II was prepared at concentrations ranging from 1-200 ppm with 5 ppm intervals. Exposure time for all solutions was 10 min. In total, 58 E. sakazakii (Cronobacter spp.) strains were tested in this study. Nine isolates were obtained from clinical samples, and 49 isolates were obtained from environmental samples. Seven strains (6 clinical and 1 environmental) were able to survive in 100 ppm disinfectant I, and a maximum of 5 ppm of disinfectant II. Fifty one strains (3 clinical and 48 environmental) were not killed in 10 ppm of disinfectant I and 1 ppm of disinfectant II in water. In conclusion, this study demonstrated that clinical E. sakazakii (Cronobacter spp.) strains displayed 5- to 10-fold higher resistance to disinfectants than environmental E. sakazakii (Cronobacter spp.) strains. Disinfectant II, containing quaternary ammonium compounds, was shown to be more potent in inactivating E. sakazakii (Cronobacter spp.) in water used to clean infant formula manufacturing equipment than disinfectant I.

Comparison of 10 Different Pre-Enrichment Broths for the Regeneration of Cronobacter spp. (Enterobacter sakazakii ) Infected in Powdered Infant Formula

  • Jung-Whan Chon;Kun-Ho Seo;Hyungsuk Oh;Dongkwan Jeong;Kwang-Young Song
    • Journal of Dairy Science and Biotechnology
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    • 제41권3호
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    • pp.103-112
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    • 2023
  • This study aimed to assess the effectiveness of 10 different pre-enrichment methods using Real-Time polymerase chain reaction (PCR) in support of the FDA method. When the initial Cronobacter spp. (Enterobacter sakazakii) inoculation was 7.2 CFU/g, the Ct values were observed in the following order: 21.37 (Enterobacteriaceae enrichment [EE] broth), 21.95 (brain heart infusion [BHI]), 22.72 (tryptic soy broth [TSB]), 23.02 (violet red bile lactose [VRBL]), 22.31 (TSB-0.1% sodium pyruvate [SP]), 23.43 (distilled water [DW]), 24.34 (phosphate buffered saline [PBS]), 24.95 (nutrient broth [NB]), 25.82 (TSB-0.6% yeast extract [YE]), and 28.27 (violet red bile glucose [VRBG]). For an inoculation of 1.82% CFU/g of Cronobacter spp. (E. sakazakii), the Ct values were recorded in this sequence: 20.34 (EE broth), 22.16 (TSB-0.6% YE), 22.37 (BHI), 22.71 (VRBL), 22.88 (TSB), 23.01 (DW), 23.19 (NB), 23.79 (TSB-0.1% SP), 24.66 (VRBG), and 24.70 (PBS). Finally, when the inoculum of Cronobacter spp. (E. sakazakii) was 0.182 CFU/g, the Ct values followed this order: 21.93 (VRBL), 23.07 (TSB-0.6% YE), 23.31 (DW), 23.47 (PBS), 23.70 (BHI), 24.14 (TSB-0.1% SP), 25.14 (TSB), 29.00 (VRBG), 31.55 (EE broth), and were undetected in the case of NB. Consequently, these results indicate that there were no significant differences among the 10 different pre-enrichment broths. Future studies should focus on exploring pre-enrichment broths that can improve the limit of detection at very low Cronobacter spp. (E. sakazakii) concentrations and enhance the selective recovery of Cronobacter spp. (E. sakazakii) under acid, antibiotic, cold, and heat damage conditions.

Cronobacter spp. 의 오염, 제어, 검출에 관한 최신 연구동향 (Current Cronobacter spp. Researches on Prevalence, Control, and Detection)

  • 송광영;천정환;김현숙;서건호
    • 미생물학회지
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    • 제48권4호
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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.

The Phenotypic and Genotypic Characterization of Korean Isolates of Cronobacter spp. (Enterobacter sakazakii)

  • Kim, Jung-Beom;Kang, Suk-Ho;Park, Yong-Bae;Choi, Jae-Ho;Park, Sung-Jin;Cho, Seung-Hak;Park, Mi-Sun;Lee, Hae-Kyung;Choi, Na-Jung;Kim, Ha-Na;Oh, Deog-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제21권5호
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    • pp.509-514
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    • 2011
  • This study was conducted to investigate the phenotypic and genotypic characteristics of Korean isolates of Cronobacter spp. (Enterobacter sakazakii). A total of 43 Cronobacter spp., including 5 clinical isolates, 34 food isolates, 2 environmental isolates, and 2 reference strains (C. sakazakii ATCC 29004 and C. muytjensii ATCC51329) were used in this study. Korean isolates of Cronobacter spp. were divided into 11 biogroups according to their biochemical profiles and 3 genomic groups based on the analysis of their 16S rRNA gene sequences. Biogroups 1 and 2 contained the majority of isolates (n=26), most of which were contained in 16S rRNA cluster 1 (n=34). Korean isolates of Cronobacter spp. showed diverse biochemical profiles. Biogroup 1 contained C. sakazakii GIHE (Gyeonggido Research Institute of Health and Environment) 1 and 2, which were isolated from babies that exhibited symptoms of Cronobacter spp. infection such as gastroenteritis, sepsis, and meningitis. Our finding revealed that Biogroup 1, C. sakazakii, is more prevalent and may be a more pathogenic biogroup than other biogroups, but the pathogenic biogroup was not represented clearly among the 11 biogroups tested in this study. Thus, all biogroups of Cronobacter spp. were recognized as pathogenic bacteria, and the absence of Cronobacter spp. in infant foods should be constantly regulated to prevent food poisoning and infection caused by Cronobacter spp.

국내 분유 및 영.유아식품에서 분리된 Cronobacter spp. (Enterobacter sakazakii)의 Biofilm 생성 특성 및 내산성 비교 (Biofilm Formation and Low pH Viability of Cronobacter spp. (Enterobacter sakazakii) Isolated from Powdered Infant Formula and Infant Foods in Korea)

  • 김선애;이유미;오세욱;곽효선;황인균;강동현;우건조;이민석
    • 한국축산식품학회지
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    • 제29권6호
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    • pp.702-708
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    • 2009
  • 본 연구는 국내에서 시판되고 있는 분유 및 영 유아식품에서 분리한 Cronobacter spp.(Enterobacter sakazakii) 14개 균주의 biofilm 생성 능력, 산성 환경에서의 생장, 용혈성을 관찰하였다. 연구 결과, 14개의 분리균주는 polyethylene에서 biofilm을 생성하였으며 영양성분이 풍부할수록 biofilm을 강하게 생성하였다. 영양성분의 종류에 따른 biofilm 생성 능력 차이는 뚜렷한 경향을 나타내지 않았으며, Cronobacter spp.의 biofilm 생성 능력은 균주별 특성에 의한 것으로 판단된다. 다양한 pH 환경에서 생장 특성을 관찰한 결과, 14개 중 7개의 균주(50%)가 pH 4.1에서도 생장할 수 있어 국외 균주에 비해 내산성이 비교적 낮게 나타났다. 한편 분리된 모든 균주에서 용혈성은 관찰되지 않았다. 본 연구는 국내 영 유아용 식품에서 분리된 Cronobacter spp.의 생리 생화학적 특성을 제시하였으며 이는 향후 Cronobacter spp. 연구 및 관련 식품산업의 저감화 기술 개발에 기초 자료로 활용될 수 있을 것으로 판단된다.

Comparison of Biochemical Profiles with Biogroups for the Identification of Cronobacter spp. (Enterobacter sakazakii)

  • Kim, Jung-Beom;Park, Yong-Bae;Kim, Woon-Ho;Kim, Ki-Cheol;Jeong, Hong-Rae;Kim, Dae-Hwan;Kang, Suk-Ho;Yong, Kum-Chan;Yoon, Mi-Hye;Park, Yong-Chul
    • 한국식품위생안전성학회지
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    • 제24권4호
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    • pp.307-311
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    • 2009
  • The objectives of this study were to compare the biochemical profiles with biogroups for the identification of Cronobacter spp. (formally known as Enterobacter sakazakii) isolates using biochemical identification kits. A total of 38 Cronobacter spp. contained 5 clinical, 31 food, and 2 environmental isolates were used. All isolates were identified as Cronobacter spp. with the Vitek II system and ID 32E kit. The API 20E kit identified all isolates as Cronobacter spp. but the percentage identification was 51.1% for 16 of 38 isolates. These strains were contained to Biogroup 2, 9, 10, and 11. The utilization of inositol is a factor determining the percentage identification of Cronobacter spp. with the API 20E kit.

Evaluation of Selective Media Containing Iron Source and Alpha-Glucosidase Substrates for Enterobacter sakazakii (Cronobacter spp.) Detection

  • Chon, Jung-Whan;Seo, Kun-Ho;Yim, Jin-Hyeok;Bae, Dongryeoul;Kim, Binn;Kim, Tae-Jin;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • 제39권1호
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    • pp.9-19
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    • 2021
  • Enterobacter sakazakii (Cronobacter spp.) causes meningitis, necrotizing enterocolitis, sepsis, and bacteremia in neonates and children and has a high mortality rate. For rapid E. sakazakii detection, various differential and selective media containing α-glucosidase substrates, such as 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside (BCIG) or 4-methylumbelliferyl-α-D-glucoside (α-MUG), have been developed as only E. sakazakii exhibits α-glucosidase activity in the genus Enterobacter. However, Escherichia vulneris (family: Enterobacteriaceae) can also utilize α-glucosidase substrates, thereby resulting in false positives. Various iron sources are known to promote the growth of gram-negative bacteria. This study aimed to develop a selective medium containing α-glucosidase substrates for E. sakazakii detection that would eliminate false positives, such as those of E. vulneris, and to determine the role of iron source in the medium. Three previously developed (TPD) media, i.e., Oxoid, OK, and VRBG, and the medium developed in this study, i.e., NGTE, were evaluated using 58 E. sakazakii and 5 non-E. sakazakii strains. Fifty-four E. sakazakii strains appeared as fluorescent or chromogenic colonies on all four media that were assessed. Two strains showed colonies on NGTE medium and not on TPD media. In contrast, the remaining two strains showed colonies on TPD media and not on NGTE medium. None of the non-E. sakazakii strains showed fluorescent or chromogenic colonies on any of the evaluated media except E. vulneris, which showed colonies on TPD media and not on NGTE medium. This study demonstrated that the newly developed NGTE medium was not only equally efficient in promoting the growth of bacterial colonies when compared with the currently available media but also eliminated false positives, such as E. vulneris.

Cronobacter Species의 검출에 관한 연구동향: 총설 (Research Trend of Cronobacter Species Detection Methods: A Review)

  • 권희준;김명희
    • 한국식품영양학회지
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    • 제28권4호
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    • pp.728-736
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    • 2015
  • Cronobacter spp.는 Salmonella spp.와 함께 조제분유에서 발견되는 미생물 중 위험도가 가장 높은 category A에 속하는 균으로 알려져 있다. 1958년 영국에서 처음으로 유아의 뇌수 막염의 원인균으로 보고되었으며, 감염 후 신생아에게서 괴사성 장염, 패혈증 등을 일으켜 후유증으로 시력과 청력 상실 및 신경마비를 일으키기도 한다. 이러한 위험성 때문에 Cronobacter spp.의 신속 검출 및 진단은 식중독 예방에 있어서 중요하다. 따라서 2002년 미국 식품의약품안전국에서는 분유에서의 Cronobacter spp.를 EE broth, VRBG 배지를 이용해 균을 분리 하고, TSA 배지에 순수분리 후 노란색을 발하는 colony를 생화학적 실험을 통해 검출하는 방법을 제시했다. 또한, 우리나라 식품공전에서도 배지를 이용하여 Cronobacter spp.를 검출 하는 방법에 대해 등재하였다. 하지만 배지배양법을 기반으로 하는 Cronobacter spp.의 검출방법은 검출에 소요되는 시간과 노동력 등이 비효율적이라는 단점이 있다. 따라서 많은 시간이 소요되는 배지배양법을 보완하고자 PCR, real-time PCR 방법 및 최근에는 PCR기법과 ELISA, CE-LIF 등을 결합하여 검출하는 방법이 개발되어 Cronobacter spp.의 검출에 사용되기도 하였다. Cronobacter spp.의 검출에 사용되고 있는 분자생물학적 기반의 PCR 및 real-time PCR 기법은 민감도와 특이도가 좋으며, 배지배양법에 의한 검출방법에 비해 검출 시간을 획기적으로 줄일 수 있다는 장점이 있다. 하지만 까다로운 실험과정과 조작에 있어서 필요한 전문적인 기술이 필요함은 한계점으로 사료된다. 면역학적 방법에 의한 Cronobacter spp. 검출에 관한 연구를 통해, 분석에 소요되는 상당시간을 단축할 수 있었으며, 민감하고 특이적인 검출이 가능함을 보여주었다. 특히 일부 연구보고에서 면역학적 검출방법은 고가의 장비가 필요 없으며, 간단한 지침에 따라 모니터링 업무의 수행이 가능하다고 하였다. 면역학적 검출방법에는 ELISA 방법 외에도 immunomagnetic bead, liposome, immunochromatographic strip 등이 개발되고 있다. 우리나라 식품의약품안전처에서는 영 유아 대상 식품의 안전관리를 강화하고자 Cronobacter spp.에 대해서는 '불검출'로 기준을 설정 운영하고 있으며, 지속적으로 이에 관한 규정을 강화하고 있는 실정이다. 따라서, 식품산업체 및 식품 의 제조, 가공, 유통 현장에서 쉽게 모니터링이 가능하며, 신속, 민감하고 특이적인 검출방법의 개발은 지속되어야 한다.

열, 살균소독제, 항생제에 의한 분리 Cronobacter spp. (Enterobacter sakazakii) 제어 (Biocontrol of Isolated Cronobacter spp. (Enterobacter sakazakii) by Heat, Sanitizer, and Antibiotic)

  • 이은진;박종현
    • 한국축산식품학회지
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    • 제30권3호
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    • pp.479-486
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    • 2010
  • 국내의 건조분말 식품에서 분리한 Cronobacter spp.의 건조내성에 따른 열, 살균소독제, 항생제에 대한 내성특성 분석하여 Cronobacter spp.을 제어하는 효과적인 방법을 연구하고자 하였다. 분리된 37개의 건조저항 균주, 7개의 건조민감 균주, 그리고 68개의 중간그룹의 균주 총 112개에 대하여 분석한 결과 건조내성을 가지고 있는 Cronobacter spp.는 열에 대해서도 저항성이 큰 것으로 나타났다. 따라서 조제분유등의 건조 스트레스에 노출되어 살아남은 Cronobacter spp.는 열 스트레스에서도 높은 내성을 보일수 있을 것으로 사료된다. 살균소독제를 이용한 Cronobacter spp.의 제어효능은 sodium hypochlorite는 15-25 ppm와 benzalkonium chloride는 5-15 ppm 농도에서 5 log CFU/mL 살균 효과가 있었다. Albumin으로 3%(w/v)오염시킨 부유세균의 살균력은 건조내성 균주와 건조민감 균주 모두에서 오염전과 비슷하여 유기물 오염상태에서도 살균소독제 효능에는 큰 영향을 주고 있지 않음을 알 수가 있었다. 그리고 항생제 내성은 $\beta$-lactam 계열의 ampicillin에 대하여 약 90% 분리주가 64-128 ppm의 농도에서 최소저해 농도를 보여 주었다. Aminoglycoside 계열의 kanamycin과 Gram(-) spectrum의 nalidixic acid에서 4 ppm의 가장 낮은 내성을 보였다. 건조민감 균주과 건조내성 균주의 최소저해농도 경우 전반적으로 모든 항생제에 대해서 건조 내성이 높은 균주가 더 큰 항생제 내성을 지니고 있는 것으로 나타났다. 따라서 분리 Cronobacter spp.의 건조내성이 열과 항생제의 저항성과 관련이 있는 것으로 보인다. 그러므로 조제분유와 같은 분말식품과 그 원료의 오염 Cronobacter spp.의 살균제어에 더 강력한 처리가 필요할 것으로 보인다.