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

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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