Sains Malaysiana 43(12)(2014): 1855–1863

 

Incidence of Cronobacter sakazakii in Powdered Infant Formula Milk Available in Malaysia

(Kehadiran Cronobacter sakazakii dalam Susu Tepung Rumusan Bayi yang Terdapat di Malaysia)

 

NORRAKIAH ABDULLAH SANI1*, MASOMEH GHASSEM1, ABDUL SALAM BABJI1,

UMA PRIYA KUPUSAMY2 & NORIZAN JAAFAR2

 

1School of Chemical Sciences and Food Technology, Faculty of Science and Technology

Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia

 

2Chemistry Department of Malaysia, 46661 Petaling Jaya, Selangor, Malaysia

 

Diserahkan: 4 Ogos 2011/Diterima: 30 September 2014

 

ABSTRACT

Enterobacter sakazakii previously known as 'yellow-pigmented E. cloacae' has been classified as a new genus 'Cronobacter'based on taxonomic analysis and geno-and phenotypic evaluation. This pathogenic organism has been associated with rare form of infant meningitis and necrotizing enterocolitis (NEC) with high mortality rate (40-80%). Some cases have been linked to the consumption of contaminated powdered infant formula milk (PIF). The objective of this study was to determine the presence of Cronobacter spp. in PIF sold in Malaysia. A selective chromogenic agar, Brilliance Enterobacter sakazakii (DFI, Oxoid), was used for detection of Cronobacter strains. Presumptive Cronobacter isolates were identified using biochemical tests (API 20E and MicrogenTM) and molecular assays (SYBR Green Real-time PCR and 16S ribosomal DNA sequencing). All presumptive Cronobacter strains produced typical blue-green colonies and non-Cronobacter strains produced yellow colonies on Brilliance Enterobacter sakazakii agar (DFI formulation). A total of 12 presumptive isolates were selected from DFI agar and identified with biochemical and molecular tests. The results indicated prevalence of 12.5% C. sakazakii contamination from 72 PIF samples. Molecular detection methods such as Real-time PCR and 16S rDNA proved to have higher identification percentage compared to the biochemical tests. In this study, it was observed that molecular assays were suitable means for sensitive identification of Cronobacter strains in PIF samples.

 

Keywords: Biochemical test; chromogenic media; Cronobacter sakazakii; enterobacteriaceae; Real time-PCR

 

ABSTRAK

Enterobacter sakazakii yang sebelum ini dikenali sebagai 'E. cloacae berpigmen kuning' telah diklasifikasikan sebagai genus baru 'Cronobacter' berdasarkan analisis takson serta semakan secara fenotip dan genotip. Organisma patogen ini dikaitkan dengan penyakit meningitis pada bayi dan enterokolitis nekrosis (NE) dengan kadar kematian yang tinggi (40-80%). Sesetengah kes dikaitkan dengan pengambilan susu tepung rumusan bayi (PIF) yang telah tercemar. Objektif kajian ini adalah untuk menentukan kehadiran Cronobacter spp. di dalam PIF di Malaysia. Suatu agar pilihan kromogenik, Brilliance Enterobacter sakazakii (DFI, Oxoid) telah digunakan bagi mengenal pasti pencilan Cronobacter. Pencilan Cronobacter jangkaan telah dikenal pasti dengan menggunakan ujian biokimia (API 20E dan MicrogenTM) dan asai molekul (SYBR Hijau Masa-nyata PCR dan penjujukan 16S DNA ribosom). Semua pencilan Cronobacter jangkaan telah menghasilkan koloni berwarna biru kehijauan manakala pencilan bukan Cronobacter menghasilkan koloni berwarna kuning di atas agar Brilliance Enterobacter sakazakii (DFI formulation). Sejumlah 12 pencilan jangkaan telah dipilih daripada agar DFI dan dikenal pasti dengan menggunakan ujian biokimia dan molecular. Hasil kajian menunjukkan kontaminasi lazim oleh C. sakazakii daripada 72 sampel PIF adalah sebanyak 12.5%. Kaedah pengesanan molekular seperti Masa-nyata PCR dan 16S rDNA telah menunjukkan peratus pengenalpastian yang lebih tinggi berbanding dengan hasil ujian biokimia. Kajian ini menunjukkan pengasaian secara molekular adalah suatu cara yang sesuai dan sensitif bagi mengenal pasti pencilan Cronobacter dalam sampel PIF.

 

Kata kunci: Cronobacter sakazakii; enterobacteriaceae; Masa-nyata PCR; medium kromogenik; ujian biokimia

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*Pengarang untuk surat-menyurat; email: norrasani@ukm.edu.my

 

 

   

 

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