Jump to ContentJump to Main Navigation
Show Summary Details
In This Section

Agricultura Tropica et Subtropica

The Journal of Czech University of Life Sciences Prague

4 Issues per year

Open Access
Online
ISSN
1801-0571
See all formats and pricing
In This Section

Isolation and characterization of Cronobacter spp. from environmental and food resources

Vannaphone Putthana
  • Corresponding author
  • Czech University of Life Sciences Prague, Institute of Tropics and Subtropics, Czech Republic
  • Email:
/ Milan Marounek
  • Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Czech Republic
/ Natalie Brenova
  • Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Czech Republic
/ Jakub Mrazek
  • Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Czech Republic
/ Daniela Lukesova
  • Czech University of Life Sciences Prague, Institute of Tropics and Subtropics, Czech Republic
Published Online: 2012-10-24 | DOI: https://doi.org/10.2478/v10295-012-0001-z

Abstract

Cronobacter spp. (formerly Enterobacter sakazakii) has been isolated from a wide range of environmental and several food sources. Cronobacter spp. is an opportunistic pathogen causing serious infection in infants, particularly neonates. The aim of this study was to isolate and characterize Cronobacter spp. from food sources (infant food, herbs and spices and vegetables) and from environmental sources as dust from vacuum cleaners. Isolation of Cronobacter spp. was performed on selective chromogenic agars, fi rstly using commercial ESIA agar and thereafter on Kim and Rhee-KR agar described in the literature. Phenotypic characteristics were obtained by commercial miniaturized biochemical ENTEROTEST 24 kits and the fi nal confi rmation of isolated strains was performed by molecular techniques (PCR, PCR - DGGE analysis, and 16S rDNA sequencing). Altogether, 99 samples were analyzed (47 samples of foods and 52 samples of dust). In total, 43 isolates of presumptive Cronobacter spp. were initially identifi ed, however, only 22 isolates (51%) were identifi ed as Cronobacter spp. with high identity scores (75-99%). The occurrence of presumptive cronobacters in environmental samples was signifi cantly higher than in samples of food (18 out of 52 vs. 4 out of 47; P = 0.003). No cronobacters were found in 17 samples of infant food, 3 isolates originated from herbs and spices, 1 isolate from spinach and 18 isolates from samples of dust (households, restaurants, dormitory rooms). It can be concluded that Cronobacter spp. is a ubiquitous pathogen contaminating food and environment. Cronobacter spp. could be well identifi ed by means of ENTERO24 test kits with high probability. Both phenotypic and genotypic methodology could be used for identifi cation of Cronobacter spp. and they can be combined for reliable identifi cation.

Keywords: : Infant food; environment; dust; Cronobacter spp.

  • BARRON J.C., FORSYTHE S.J. (2007): Dry stress and survival time of Enterobacter sakazakii and other Enterobacteriaceae in dehydrated powdered infant formula. Journal of Food Protection 70: 2111-2117.

  • BESSE N.G., LECLERCQ A., MALADEN V., TYBURSKI C., BERTRAND L. (2006): Evaluation of the international organization for standardization-International Dairy Federation (ISO-IDF) draft standard method for detection of Enterobacter sakazakii in powdered infant food formulas. Journal of AOAC (Association of Offi cial Analytical Chemists) International 89 (5):1309-1316

  • ČSN P ISO/TS 22964. (2006): Milk and milk products- Detection of Enterobacter sakazakii. Czech Standards Institute.

  • FARMER J.J., ASBURY M.A., HICKMAN F.W., BRENNER D.J. (1980): Enterobacter sakazakii: A new species of Enterobacteriaceae isolated from clinical specimens. International Journal of Systematic Bacteriology 30: 569 - 584. [Crossref]

  • FORSYTHE S.J. (2005): Enterobacter sakazakii and other bacteria in powdered infant milk formula. Maternal Child Nutrition 1(1): 44-50.

  • FORSYTHE S. J. (2010): Cronobacter species. Culture 31(1): 1-8. Ed.: Oxoid Ltd. ISSN 0965-0989

  • FRIEDEMANN M. (2007): Enterobacter sakazakii in food and beverages (other than infant formula and milk powder). International Journal of Food Microbiology 116: 1-10. [Web of Science]

  • GURTLER J.B., KORNACKI J.L., BEUCHAT L.R. (2005): Enterobacter sakazakii: A coliform of increased concern to infant health. International Journal of Food Microbiology 104: 1-34.

  • HASSAN A.A., AKINEDEN O., KRESS C., ESTUNINGSIH S., SCHNEIDER E., USLEBER E. (2007): Characterization of the gene encoding the 16S rRNA of Enterobactersakazakii and development of a species-specifi c PCR method. International Journal of Food Microbiology 116: 214-220.

  • IVERSEN C., FORSYTHE S.J. (2004): Isolation of Enterobactersakazakii and other Enterobacteriaceae from powdered infant milk and related products. International Journal of Food Microbiology 21: 771-777.

  • IVERSEN C., FORSYTHE S. (2003): Risk profi le of Enterobacter sakazakii, an emergent pathogen associated with infant milk formula. Trends in Food Science Technology 14: 443-454. [Crossref]

  • IVERSEN C., DALE H., DRUGGAN P., WAD DINGTON M., CRAWFORD J., FORSYTHE S.J. (2004): The identifi cation of Enterobacter sakazakii using partial 16S sequencing and biochemical techniques. 104th ASM General Meeting, New Orleans, LA.

  • IVERSEN C., LEHNER A., MULLANE N., BIDLAS E., CLEENWERCK I., MARUGG J., FANNING S., STEPHAN R., JOOSTEN H. (2007): The taxonomy of Enterobactersakazakii: proposal of a new genus comb. nov. Cronobacter sakazakii subsp. sakazakii, comb. nov., Cronobacter sakazakii subsp. malonaticus subsp. nov., Cronobacter turicensis sp. nov., Cronobactermuytjensii sp. nov., Cronobacter dublinensis sp. nov. and Cronobacter genomospecies 1. BioMed Central Evolutionary Biology 7: 64.

  • IVERSEN C., MULLENE M., MCCARDELL B., TALL B.D., LEHNER A., FANNING S., STEPHAN R., JOOSTEN H. (2008): Cronobacter gen. nov., a new genus to accommodate the biogroups of Enterobacter sakazakii, and proposal of Cronobacter sakazakii gen. nov., comb. nov., C. malonaticus sp. nov., C. turicensis, sp. nov., C. muytjensii sp. nov., C. dublinensis sp. nov., Cronobacter genomospecies 1, and of three subspecies. C. dublinensis sp. nov. subsp. Dublinensis subsp. nov. C. dublinensis sp. nov. subsp. lausannensis subsp. nov., and C. dublinensis sp. nov. subsp. lactaridi subsp. nov. International Journal of Systematic and Evolutionary Microbiology 58: 1442-1447.

  • JARADAT Z.W., ABABNEH Q.O., SAADOUN I.M., SAMARA N.A., RASHDAN A.M. (2009): Isolation of Cronobacter spp. (formerly Enterobacter sakazakii) from infant food, herbs and environmental samples and the subsequent identifi cation and confi rmation of the isolates using biochemical, chromogenic assays, PCR and 16S rRNA sequencing. BioMed Central Microbiology 9: 225.

  • KANDHAI M.C., REIJ M.W., GORRIS L.G.M., GUILLAUMEGENTIL O., VAN SCHOTHORST M. (2004): Occurrence of Enterobacter sakazakii in food production environments and households. Lancet 363: 39-40.

  • KIM S.A., RHEE M.S. (2011): A new cost-effective, selective and differential medium for isolation of Cronobacter spp. Journal of Microbiology Methods 85: 149-154. [Web of Science]

  • KUCEROVA E., CLIFTON S. W., XIAO-QIN X., LONG F., PORWOLLIK S., FULTON L., FRONICK C., MINX P., KY- UNG K., WARREN W., FULTON R., FENG D., WOLLAM A., SHAH N., BHONAGIRI V., NASH W.E., HALLSWORTH-PEPIN K., WILSON R.K., MCCLELLAND M., FORSYTHE S.J. (2010): Genome Sequence of Cronobacter sakazakii BAA- 894 and Comparative Genomic Hybridization Analysis with Other Cronobacter Species. PLoS ONE 5 (3): e9556.

  • LAI K.K. (2001): Enterobacter sakazakii infections among neonates, infants, children, and adults. Case reports and a review of the literature. Medicine 80: 113-122.

  • LEHNER A., NITZSCHE S., BREEUWER P., DIEP B., THELEN K., STEPHAN R. (2006): Comparison of two chromogenic media and evaluation of two molecular based identifi cation systems for Enterobacter sakazakii detection. BioMed Central Microbiology 6: 15

  • MUYZER G., DE WAA, E.C., UITIERLIN DEN A.G. (1993): Profi ling of Complex Microbial Populations by Denaturing Gradient Gel Electrophoresis Analysis of Polymerase Chain Reaction-Amplifi ed Genes Coding for 16S rRNA. Applied Environmental Microbiology 59: 695-700.

  • MUYTJENS H.L., VAN DER ROS-VAN DER REPE J., VAN DRUTEN H.A.M. (1984): Enzymatic profi les of Enterobacter sakazakii and related species with special reference to the alpha-glucosidase reaction and reproducibility of the test system. Journal of Clinical Microbiology 20: 684-686.

  • OH S.W., KANG, D.H. (2011): Fluorogenic selective and differential medium for isolation of Enterobacter sakazakii. Applied Environmental Microbiology 70: 5692-5694.

  • RESTINO L., FRAMPTON E.W., LIONBERG W.C., BECKER R.J. (2006): A chromogenic plating medium for the isolation and identifi cation of Enterobacter sakazakii from foods, food ingredients and environmental sources. Journal of Food Protection 69: 315-322.

  • SIMMONS B.P., GELFAND M.S., HAAS M., METTS L., FERUSON J. (1989): Enterobacter sakazakii infection in neonates associated with intrinsic contamination of powdered infant formula. Infection Control and Hospital Epidemiology 10: 398-401. [Crossref]

  • STOOP B., LEHNER A., IVERSEN C., FANNING S., STEPHAN R. (2009): Development and evaluation of rpoB based PCR systems to differentiate the six proposed species within the genus Cronobacter. International Journal of Food Microbiology 136: 165-168. [Web of Science]

  • URMENYI A.M.C., FRANKLIN A.W. (1961): Neonatal death from pigmented coliform infection. Lancet 277: 313- 315. Received for publication: December 21, 2011Accepted for publication: February 23, 2012

About the article

Published Online: 2012-10-24

Published in Print: 2012-03-01



Citation Information: Agricultura tropica et subtropica, ISSN (Online) 1801-0571, DOI: https://doi.org/10.2478/v10295-012-0001-z. Export Citation

This content is open access.

Comments (0)

Please log in or register to comment.
Log in