Investigation of Extended Spectrum Β-Lactamases (ESBL)-Producing Enterobacteriaceae and Cronobacter Spp in Infant Formulas and Cereal-Based Foods for Children

Year 2017, Issue: 1, 19 - 32, 26.04.2017

Pınar Cava Gümüş İsmail Hakkı Tekiner Burcu Çakmak Zeynep Tacer Caba Haydar Özpınar

Abstract

Aim: The
extended spectrum β-lactamase (ESBL) producing Enterobacteriaceae have attracted attention due to their ability of
delaying or cancelling antimicrobial therapy. The foods for infants and
children may occasionally contain pathogens such as ESBL-producing Enterobacteriaceae, including Cronobacter spp., because of
inappropriate production, storage and handling conditions. This situation leads
to a greatest risk of infection to infants and children through their consumption.
The objective of this study was to investigate the occurrence of ESBL-producing
Enterobacteriaceae, including Cronobacter spp., from a total of 115
samples of various foods from different brands for infants and children sold in
Istanbul, Turkey (20 locally produced infant formula, 20 imported infant
formula, 20 starch, 20 rice flour, 20 semolina, and 15 milk powder).

Methods: For isolation of Enterobacteriaceae ISO 21528-2:2004 was
followed. The isolated colonies were identified by Mass Spectrometer. The identified
colonies were then exposed to pre-enrichment at 35-370C for 18-24
hours, the suspension was transferred to a Chromogenic ESBL selective media,
and allowed for an incubation at 35-370C for 18-24 hours again. The
presumptive ESBL producers were subjected to ESBL screening test using a
combination of ceftazidime, cefotaxime, and cefpodoxime ± clavulanic acid.
Finally, MIC values for ESBL confirmation were conducted by Merlin Micronaut-S
beta-lactamase VII kit, the readings were automatically evaluated by Sifin
Software integrated with a spectrometer. All procedures were performed
according to the Guidelines of CLSI (2013).

Results: Of 18 ESBL suspected isolates (7 P. agglomerans, 1 E. aerogenes, 3 E.  cloacae/asburiae, 2 K. oxytoca, 2 L. adecarboxylata,
and 3 C. sakazakii), 5 isolates (3 P. agglomerans, 1 E. cloacae/ asburiae, and 1 E.
aerogenes) were positive for ESBL-production, whereas none of C. sakazakii isolates was
ESBL-producers.

Discussion and Conclusion: Some of the
analysed infant formulas and cereal-based foods for children (locally produced
infant formulas, rice flour, and starch) were determined to be posing a serious
health risk for the infants and children due to contamination by ESBL-producing
Enterobacteriaceae, except for C. sakazakii isolates.

Keywords

Cereal-based food, Children, Cronobacter spp., Enterobacteriaceae, ESBL, Infant formula

Bebek Mamaları ve Tahıl Bazlı Çocuk Gıdalarında Genişlemiş Spektrumlu Β-Laktamazlar (GSBL) Üreten Enterobacteriaceae ve Cronobacter Spp. Araştırılması

Abstract

Amaç: Genişlemiş spektrumlu β-laktamaz
(GSBL) üreten Enterobacteriaceae türleri
antimikrobiyal tedavileri başarısız kıldıkları için artan şekilde ilgi
görmektedirler. Yenidoğan ve çocuk gıdaları uygunsuz üretim, depolama ve
hazırlama koşulları sebebiyle GSBL-üreten Enterobacteriaceae
suşları, özellikle Cronobacter
türleri dahil olmak üzere, patojen mikroorganizmalar içerebilmektedir. Bu tür
gıdaların tüketilmesi yenidoğan ve çocuk sağlığı açısından son derece tehlikeli
enfeksiyonlara yol açmaktadır. Bu çalışmada İstanbul ilinde satışa sunulan
farklı markalara ait toplam 115 örnekte (20 yerli üretim bebek maması, 20 ithal
bebek maması, 20 nişasta, 20 pirinç unu, 20 irmik ve 15 süt tozu) GSBL-üreten Enterobacteriaceae ve Cronobacter türlerinin varlıklarının
araştırılması amaçlanmıştır.

Yöntem: Örneklerde Enterobacteriaceae türlerinin izolasyonu ISO 21528-2:2004
talimatları takip edilerek yapılmıştır. İzole edilen Enterobacteriaceae türleri Kütle Spektrometresi ile
tiplendirilmiştir. Tiplendirmesi tamamlanmış izolatlar ön zenginleştirme amaçlı
35-370C/18-24 saat inkübe edildikten sonra, kromojenik GSBL seçici
besiyerine transfer edilmiş ve tekrar 35-370C/18-24 saat inkübasyona
bırakılmıştır. İnkübasyon sonunda gelişen GSBL şüpheli koloniler seftazidim,
sefotaksim ve sefpodoksim ± klavulanik asit içeren antibiyotik diskler
kullanılarak incelenmiştir. İnceleme sonucu güçlü şüpheli GSBL pozitif olduğu
belirlenen kolonilerin MİK değeri tespiti Merlin Micronaut-S beta-lactamase VII
kiti kullanılarak yapılmış, MİK bulguları spektrometre tarafından ölçülmüş ve
Sifin yazılımı ile otomatik olarak değerlendirilmiştir. GSBL tarama ve MİK
tespiti CLSI (2013) talimatları takip edilerek yapılmıştır.

Bulgular: Mikrobiyolojik İnceleme sonucu elde
edilen toplam 18 adet GSBL şüpheli izolat arasında (7 adet P.  agglomerans, 1 adet E. aerogenes, 3 adet E. 
cloacae/asburiae, 2 adet K.
oxytoca, 2 adet L. adecarboxylata
ve 3 adet C. sakazakii), 5 adet
izolatın (3 adet P. agglomerans, 1
adet E. cloacae/ asburiae ve 1 adet E. aerogenes) GSBL pozitif oldukları,
diğer taraftan C. sakazakii
izolatların GSBL üretmedikleri tespit edilmiştir.

Tartışma ve Sonuç: Yerli üretim
bebek mamaları, pirinç unu ve nişasta örneklerinin, C. sakazakii izolatları hariç, GSBL-üreten Enterobacteriaceae türleri içerdikleri, bu sebeple bebek ve çocuk
sağlığı açısından ciddi sağlık riski oluşturdukları görülmüştür.

Keywords

Tahıl bazlı gıda, Çocuk, Cronobacter spp., Enterobacteriaceae, GSBL, Bebek maması

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