Ketahanan dan Kulturabilitas Cronobacter Sakazakii terhadap Stres Kering pada Simulasi Proses Pengeringan

  • Siti Nurjanah 1) Departemen lImu dan Teknologi Pangan, Fakultas Teknologi Pertanian, lnstitut Pertanian Bogor, Bogor dan 2) South East Asian Food and Agricultural Science and Technology Center, Institut Pertanian Bogor, Bogor
  • Ratna Nurmalita Sari Departemen lImu dan Teknologi Pangan, Fakultas Teknologi Pertanian, lnstitut Pertanian Bogor, Bogor
  • Ratih Dewanti-Hariyadi 1) Departemen lImu dan Teknologi Pangan, Fakultas Teknologi Pertanian, lnstitut Pertanian Bogor, Bogor dan 2) South East Asian Food and Agricultural Science and Technology Center, Institut Pertanian Bogor, Bogor
Keywords: Cronobacter sakazakii, culturability, drying, resuscitation, survival


Cronobacter sakazakii is an opportunistic pathogen that has been isolated from several dried food products and reported can survive in dry conditions. Survival on dry stress can be observed using microscopy due to losses of their culturability in the medium. This study aimed to observe the survival and culturability of C. sakazakii on dry stress and ability to recover their culturability or resuscitation. This research was carried out in 3 steps: drying cells by desiccation, survival cell enumeration, and resuscitation. The bacteria used were two isolates Green fluorescence protein (pGFPuv) mutants of C. sakazakii, namely E2 and Yrt2a, that have ampicillin resistance and similar growth pattern with their wild-type. Desiccation was conducted by placed bacterial cells in incubator at 30, 35, 400 and 500C for 2 hours and air-drying stored at 210C for 72 hours. The culture able cells were enumerated on tryptone soy agar (TSA) and total cells include non-culturable cells were enumerated using fluorescence microscopy. Both of C. sakazakii isolates can survive for all of treatment. Total cells of E2 and Yrt2a decreased 2-3 and 4-5 log cells respectively. Both of isolates loss their ability to grow on medium, however suplementation the medium growth using pyruvate can resuscitate their cells and recover their culturability. Resuscitation of C. sakazakii cells might be as potential risk for increasing bacterial contaminant in food.


Download data is not yet available.


[BAM]. 2001. Chapter 3: Aerobic plate count. [5 Desember 2015]

[WHO]. 2008. Enterobacter sakazakii and Other Microorganism in Powdered Infant Formula. Microbiol Risk Asses. WHO, Geneva, Switzerland.

Besnard V, Frederigh M, Declerq E, Jugiau F, Ceppelier J. 2002. Environmenta and physico-chemical factors induce VBNC state Listeria monocytogenes. J Vet Res 33: 395 – 370. DOI: 10.1051/vetres:2002022.

Beuchat L, Kim H, Gutler J, Lin L, Ryu J, Glenner, Richards. 2009. Cronobacter sakazakii in foods and factors affecting its survival, growth, and inactivation. J Food Microbiol 136: 204 – 213. DOI: 10.1016/j.ijfoodmicro.2009.02.029.

Breeuwer P. Lardeau A, Joosten H. 2003. Dessication and heat tolerance of Enterobacter sakazakii. J. Appl Microbiol 95: 967–973. DOI: 10.1046/j.1365-2672.2003.02067.x.

Dancer G, Mah J, Rhee S, Hwang I, Kang D. 2009. Resistence of Enterobacter sakazakii to environ-mental stresses. J Appl Microbiol 107: 1606–1614. DOI: 10.1111/j.1365-2672.2009.04347.x.

Dewanti-Hariyadi R, Gitapratiwi D, Meutia Y, Hidayat S, Nurjanah S. 2010. Isolation of Cronobacter sakazakii from Powdered Infant Formula and Other Dried Food Obtained from Bogor in: International Seminar on Current Issues Challenges in Food Safety: Science-Based Approach for Food Safety Management. October 3–4, IPB International Confrence Center, Bogor, Indonesia. South Asian Food and Agricultural Science and Technology (SEAFAST) Center, Bogor, Indonesia.

Dewanti-Hariyadi R, Larasati F, Nuraida L. 2012. Survival of Cronobacter sakazakii in skim milk during spray drying storage and resuscitation. J Teknol Industri Pangan 13: 186-192. DOI: 10.6066/ jtip.2012.23.2.186.

Downing K, Mischenko V, Shleeva, Young D, Young M, Kaprelyants A. 2005. Mutants of Myco-bacterium tuberculosis lacking three of the five rpf-like gens are defective for growth in vivo and resuscitation in vitro. J Infect Immun 73: 3038–3043. DOI: 10.1128/IAI.73.5.3038-3043.2005.

Du M, Chen J, Zhang X, Li A, Wang Y. 2007. Retention of virulence in a viable but nonculturable Edwardsiella tarda isolate. Appl Environ Microbiol 73: 1349–1354. DOI: 10.1128/AEM.02243-06.

Ducret A, Chablier M, Dukan S. 2014. Characterization and resuscitation of non-culturable cells of Legionela pneumophilla. BMC Microbol 14: 3. DOI: 10.1186/1471-2180-14-3.

Estuningsih S, Kress C Hassan A, Akinden O, Schineider E, Usleber E. 2006 Cronobactericeae in dehydrated powdered infant formula manufactured in Indonesia and Malaysia. J Food Prot 69(12): 3013–3017. DOI: 10.4315/0362-028X-69.12.3013.

Friedmann M. 2007. Enterobacter sakazakii in food and beverages (other than infant formula and milk powder). Int J Food Microbiol 116: 1–10. DOI: 10.1016/j.ijfoodmicro.2006.12.018.

Gitapratiwai D, Dewanti-Hariyadi R, Hidayat SH. 2012. Genetic relatedness of Cronobacter spp isolated from dried food product in Indonesia. J Int Food Res 17(4): 1745-1749.

Haddix P, Paulsen E, Werner T. 2000. Measurment of mutation to antibiotic resistance: ampicilin resistance in Serratia marcescens: J Bioscene 26 (1): 17–21.

Iversen C, Forsythe S. 2004. Isolation Enterobacter sakazakii and other Enteroobacteriaceae from powdered infant milk and related products. J Food Microbiol 21: 771–777. DOI: 10.1016/ 01.009.

Li L, Mendis N, Trigui H, Oliver J, Faucher S. 2014. The importance of the viable but non-culturable state in human bacterial pathogens. Frontiers Microbiol 5(258): 1–20. DOI: 10.3389/fmicb.2014. 00258.

Listyoningrum H, Harijono. 2015. Optimasi susu bubuk dalam makanan pendamping ASI. J Pang Agr 3(4): 1302–1312.

Liu Y, Gilchrist A, Zhang J, Li F. 2008. Detection of viable but non-culturable Eschericia coli O157;H7 bacteria in drinking water and river water. Appl Environ Microbiol 74(5): 1502–1507. DOI: 10.11 28/AEM.02125-07.

Lleo M, Tafi M, Canepari P. 1998. Nonculturable Enterococus faecalis cells are metabolically active and capable of resuming active growts. J Syst Appl Microbiol 21: 333–339. DOI: 10.1016/S0723-2020(98)80041-6.

Lleo M, Benedetti, Tafi D, Signoretto C, Canepari P. 2007. Inhibition of resuscitation from the viable but non-culturable state in Enterococcus faecalis. J Environ Microbiol 9: 2313–2320. DOI: 10.1111/j. 1462-2920.2007.01345.x.

Majumdar SA, Chen XD. 2009. Drying Technologies in Food Processing. New Jersey (US): Blackwell Publishing.

Meutia YR, Dewanti R, Estuningsih S. 2008. Enterobacter sakazakii Isolate Asal Susu Formula dan Makanan Bayi: Karakterisasi Gen 16s rRNA dan Perilaku Bakteri Pasca Rekonstitusi. [Desertasi]. Bogor, IPB.

Mizunoe Y, Wai A, Takade A, Yoshida S. 2000. Resuscitation of viable but nonculturable cells of Vibrio parahaemolyticus induced at low tempe-rature under starvation. FEM Microbiol Letters 186(2000): 115–120. DOI: 10.1111/j.1574-6968.2000.tb09091.x.

Musa AMA. 2015. Using pGFP Mutants to Study the Influence of Drying on the Survival of Cronobacter sakazakii in Maize. [Thesis]. Sekolah Pascasarjana, IPB, Bogor.

Nabulsi A, Osaili T, Al-Holy M, Shaker R, Ayyash M, Olaimat A, Holley R. 2009. Influence of dessication on the sensivity of Cronobacter spp. to lactoferrin ornisin in broth and powdered infant formula. Int J Food Microbiol 136: 221–226. DOI: 10.1016/j.ij foodmicro.2009.08.008.

Nurjanah S. 2014. Sitotoksisitas dan Pelabelan Cronobacter sakazakii dengan Green Fluorescent Protein untuk Mempelajari Perilakunya selama Pengeringan Jagung. [Desertasi]. Sekolah Pasca-sarjana, IPB, Bogor.

Oliver J. 2005. The viable but nonculurable state in bacteria. J Microbiol 43(1): 93–100.

Pederson J, Jacobsen C. 1993. Fate of Enterobacter cloacae JP210 and AEO106 (pRo101) in soil during waterstress: Effect on culturability and viability. J Appl Environ Microbiol 59(5): 1560–1564.

Pinto D, Santos M, Chambel. 2013. Thirty years of viable but nonculturable state research: Unsolved molecular mechanism. Crit Rev Microbiol 41(1): 61-76. DOI: 10.3109/1040841X.2013.794127.

Potts M. 1994. Desiccation tolerance of prokaryotes. J Microbial Rev 58: 755 – 805.

Sally C, Dickson J. 2004. Survival and recovery of viable but non-culturable Listeria monocytogenes cells in a nutritionally depletion. J Food Prot 67(8): 1641–1645. DOI: 10.4315/0362-028X-67.8.1641.

Sardessai Y. 2005. The viable but nonculturable bacteria: their impact on public health. J Curr Sci 89(10): 1650.

Shaker R, Osaili A, Hasan A, Ayyash M, Forsythe S. 2008. Effect of desiccation, starvation, heat, and cold stresses on the thermal resistance of Enterobacter sakazakii in rehydrated infant milk formula. J Food Sci 73(7): 354–359. DOI: 10.11 11/j.1750-3841.2008.00880.x.

Silitonga YW. 2016. Resistensi Cronobacter sakazakii terhadap Ampisilin dan Hubungannya dengan Stabilitas Ekspresi GFPuv. [Thesis]. Sekolah Pascasarjana, IPB, Bogor.

Sulistiyanti ST. Kajian Pembuatan Maizena dari Jagung Kuning dan Sintas Mutan Cronobacter spp. Selama Pembuatan Maizena. [Skripsi]. Fakultas Teknologi Pertanian, IPB, Bogor.

Theresia R. 2015. Ketahanan mutan Cronobacter sakazakii dan Mikroorganisme lain terhadap Ampisilin untuk Pengembangan Media Spesifik. [Skripsi]. Fakultas Teknologi Pertanian, IPB, Bogor.

Vriezen J, Bruijn F, Nusslein K. 2012. Dessication induces viable but non-culturable cells in Sinorhizobium meliloti 1021. AMB Express 2(6): 1–9. DOI: 10.1186/2191-0855-2-6.

Yousef AE, Carlstrom C. 2003. Food Microbiology A Laboratory Manual. New Jersey: John Wiley & Sons.

How to Cite
NurjanahS., SariR. N., & Dewanti-HariyadiR. (2017). Ketahanan dan Kulturabilitas Cronobacter Sakazakii terhadap Stres Kering pada Simulasi Proses Pengeringan. Jurnal Mutu Pangan : Indonesian Journal of Food Quality, 4(2), 92-99. Retrieved from
Research Paper