Isolasi dan Identifikasi Enterobacter sakazakii pada Susu Mentah dan Produk Susu Segar di Daerah Istimewa Yogyakarta

Tri Yahya Budiarso; Hutri Catur Sad Winarni

doi:10.22146/jsv.27565

Isolasi dan Identifikasi Enterobacter sakazakii pada Susu Mentah dan Produk Susu Segar di Daerah Istimewa Yogyakarta

https://doi.org/10.22146/jsv.27565

Tri Yahya Budiarso^(1*), Hutri Catur Sad Winarni⁽²⁾

(1) Fakultas Bioteknologi, Universitas Kristen Duta Wacana
(2) Fakultas Bioteknologi, Universitas Kristen Duta Wacana
(*) Corresponding Author

Abstract

Enterobacter sakazakii is an opportunistic pathogen that causes infections such as necrotizing enterocolitis, bacteremia, and meningitis which, in many cases, have been associated with the consumption of powdered infant formula. Raw milk has been considered to be a potential source of contamination. The aim of
this study was to detect and identify the presence of E. sakazakii in raw milk from local farms and fresh dairy products sold by street vendors and cafés in the Special Region of Yogyakarta. A total of 80 milk samples were cultured in Enterobacterriaceae Enrichment Broth. The cell cultures were then enumerated on Tryptone Soya Agar and Chromocult Coliform Agar. For the isolation of suspect colonies, Chromogenic Druggan-Forsythe-Iversen (DFI) Agar was used. Presumptive colonies of E.sakazakii were characterized biochemically using API
20E and molecularly through the amplification of the 16S rRNA gene using primer pairs 16_SUNI-L/Saka-2b and ESA-1/16_SUNI-R. Based on the α-glucosidase test of DFI Agar, only 9 isolates out of the 80 samples were
identified as suspected E.sakazakii. Biochemical and molecular identification suggested that 5 isolates resulted positive for E.sakazakii, while the other 4 were Panthoea spp1.

Keywords

isolation; identification; E.sakazakii; raw milk; fresh milk

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References

Chen, Y., Song, K.-Y., Brown, E. W., & Lampel, K. A.

(2010) Development of an improved protocol for the isolation and detection of Enterobacter sakazakii (Cronobacter) from powdered infant formula, J. of Food Protec. 73: 1016–1022.

Desty, G., Dewanti Haryadi, R., Hidayat,S.H. (2012)

Genetic relatedness of Cronobacter spp.

(Enterobacter sakazakii) isolated from dried

food products in Indonesia, Int. Food Research

Journal 19: 1745–1749.

Drudy, D., Wall, P. G., Mullane, N. R., Quinn, T.,

Wall, P. G., & Fanning, S. (2006) Enterobacter

sakazakii : An Emerging Pathogen in Powdered

Infant Formula, Food Safety CID 2006: 42996-

http://doi.org/10.1086/501019.

Farmer III, J.J., Asbury, M.A., Hickman, F.W.

Brenner, D.J. (1980) Enterobacter sakazakii:

A New Species of “ Enterobacteriaceae ”

Isolated from Clinical Specimens, Int. J.of

Syst.Bact. 30: 569-584.

FDA. (2002) Food Isolation and Enumeration of

Enterobacter sakazakii from Dehydrated

Powdered Infant Formula, http://www.

fda.gov/Food/FoodScienceResearch/

LaboratoryMethods/ucm114665. htm.

Hassan, A. A., Akineden, Ö., Kress, C., Estuningsih,

S., Schneider, E., & Usleber, E. (2007)

Characterization of the gene encoding the

S rRNA of Enterobacter sakazakii and

development of a species-specific PCR

method, I.J.Food.Micro. 116: 214–220. http://

doi.org/10.1016/j.ijfoodmicro. 2006.12.011.

Haug, A., Høstmark, A. T., & Harstad, O. M. (2007)

Bovine milk in human nutrition a review.

Lipids in Health and Disease, BMC 6:25.

http://doi.org/10.1186/1476-511X-6-25.

Hill, B., Smythe, B., Lindsay, D., & Shepherd, J.

(2012) Microbiology of raw milk in New

Zealand, I.J.Food.Micro, 157: 305–308. http://

doi.org/10.1016/j.ijfoodmicro.2012.03.031.

Iversen, C., Druggan, P., & Forsythe, S. (2004) A

selective differential medium for Enterobacter

sakazakii, a preliminary study, I.J.Food.

Micro. 96: 133–9. http://doi.org/10.1016/

j.ijfoodmicro. 2004.01.024.

Iversen, C., & Forsythe, S. (2004) Isolation

of Enterobacter sakazakii and other

Enterobacteriaceae from powdered infant

formula milk and related products, Food

Micro 21: 771–777. http://doi.org/10.1016/j.

fm.2004.01.009.

Iversen, C., & Forsythe, S. J. (2007) Comparison

of media for the Isolation of Enterobacter

sakazakii, AEM 73: 48–52. http://doi.

org/10.1128/AEM.01562-06.

Iversen, C., Lehner, A., Mullane, N., Bidlas, E.,

Cleenwerck, I., Marugg, J.,..Joosten., H.

(2007) The taxonomy of Enterobacter

sakazakii: proposal of a new genus Cronobacter

gen. nov. and descriptions of Cronobacter

sakazakii comb. nov. Cronobacter sakazakii

subsp. sakazakii, comb. nov., Cronobacter

sakazakii subsp. malonaticus subsp. nov.,

Cronobacter turicensis sp. nov., Cronobacter

muytjensii sp. nov., Cronobacter dublinensis

sp. nov. and Cronobacter genomospecies 1,

BMC Evollutionaryiology 7: 64. http://www.

biomedcentral.com/1471-2148/7/64.

Iversen, C., Mullane, N., McCardell, B., Tall, B.

D., Lehner, A., Fanning, S., ..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., Cronobacter

malonaticus sp. nov., Cronobacter turicensis

sp. nov., Cronobacter muytjensii sp. nov.,

Cronobacter dublinensis sp. nov., Cronobacter

genomo species 1, and of three subspecies,

Cronobacter dublinensis subsp. dublinensis

subsp. nov., Cronobacter dublinensis subsp.

lausannensis subsp. nov. and Cronobacter

dublinensis subsp. lactaridi subsp. nov., I.J.of

Syst. and Evolutionary Micro, 58: 1442–1447.

http://doi.org/10.1099/ijs.0.65577-0.

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

identification and confirmation of the isolates

using biochemical, chromogenic assays, PCR

and 16S rRNA sequencing, BMC Micro. 9:225.

http://doi.org/10.1186/1471-2180-9-225.

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 identification systems for

Enterobacter sakazakii detection, BMC Micro.

: 15. http://doi.org/10.1186/1471-2180-6-15.

Leuschner, R. G. K., Baird, F., Donald, B., & Cox, L. J. (2004) A medium for the presumptive detection of Enterobacter sakazakii in infant formula, Food Micro. 21: 527–533. http://doi.org/10.1016/j.fm. 2003.12.002

Lou, X., Si, G., Yu, H., Qi, J., Liu, T., & Fang, Z. (2014)

Possible reservoir and routes of transmission of Cronobacter (Enterobacter sakazakii) via wheat flour, Food Control 43: 258–262. http://doi.org/10.1016/j.foodcont.2014.03.029.

Manafi, M. (2003) Media for detection and enumeration of “total” Enterobacteriaceae, coliforms and Escherichia coli from water

and foods, Handbook of Culture Media for Food Microbiology p:167–193. http://doi.org/10.1016/S0079-6352(03)80015-2.

Muytjens, H. L., J. van Spaceder Ros-van de Repeqq,and H. A. M. van Druten. (1984) Enzymatic Profiles of Enterobacter sakazakii and

Related Species with Special Reference to the α-Glucosidase Reaction and Reproducibility of the Test System, J. Clin. Microbiol. 20:684–686.

Rattanabumrung, O., Sangadkit, V., Supanivatin, P., & Thipayarat, A. (2012) Kinetics of E.coli colony area expansion and color

development in Chromocult Coliform Agar (CCA) under different incubation conditions, Procedia Engineering 32: 134–140. http://doi.

org/10.1016/j.proeng.2012.01.1247.

Yan, Q. Q., Condell, O., Power, K., Butler, F., Tall, B.D., & Fanning, S. (2012) Cronobacter species (formerly known as Enterobacter sakazakii) in powdered infant formula: a review of our current understanding of the biology of this bacterium, J. of Applied Micro. 113:1–15. http://doi.org/10.1111/j.1365-2672.2012.05281.x

DOI: https://doi.org/10.22146/jsv.27565