RESUMO: Dentre o gênero Cronobacter, Cronobacter sakazakii é a espécie mais comum que representa um grave risco para a saúde dos recém-nascidos, bebês e crianças. Algumas formulas infantis paracrianças, alimentos a base de cereais e locais de produção de alimentos, foram considerados como potenciais locais de contaminação de C. sakazakii. Este patógeno possui diferentes agentes de virulência codificados por diferentes genes de virulência. Portanto, a caracterização dos genes é importante para distinguir as cepas patogênicas das não patogênicas. O objetivo deste estudo foi caracterizar os diferentes genes de virulência [OmpA, OmpX, zpx, and Cpa] em C. sakazakii isolados em um total de 120 amostras (20 de cada uma delas - leite em pó, amido, farinha de arroz, sêmola, comida para bebês e amostras de poeira provenientes dos ambientes de produção de alimentos) por reação em cadeia da polimerase em tempo real (RT-PCR). No total, foram cultivadas 13 estirpes (7 de leite em pó, 2 de farinha de arroz, 1 de sêmola e 3 poeiras) e depois identificadas pelo kit de teste bioMérieux API® 20E. As estirpes identificadas foram submetidas ao PCR em tempo real para caracterizar os genes alvo associados à virulência. Os resultados mostraram que todos os genes C. sakazakii isolados eram patogenicos e positivos às OmpA, OmpX, zpx e Cpa com um padrão de coexistência. Em resumo, o nosso estudo revelou que os alimentos e os ambientes de produção de alimentos analisados constituíam uma ameaça à saúde dos recém-nascidos, bebês e crianças devido à contaminação por C. sakazakii patogênico como marcadores associados à virulência.

ABSTRACT: Among the Cronobacter genus, Cronobacter sakazakii is the most common species posing a severe health risk for newborns, infants and children. Some infant formulas, cereal-based foods, and food production environments may be the potential reservoirs of C. sakazakii. This pathogen possesses different virulence factors encoded by different virulence genes. Therefore, characterizing these genes is important for distinguishing pathogenic strains from nonpathogenic ones. The objective of this study was to characterize some virulence genes [OmpA, OmpX, zpx, and Cpa] by real-time polymerase chain reaction (PCR) in C. sakazakii isolates from a total of 120 samples (20 each of milk powder, starch, rice flour, semolina, infant formula and dust samples from food production environments). Overall, 13 isolates (7 from milk powder, 2 rice flour, 1 semolina, and 3 dust) were cultured, identified by bioMérieux API® 20E test kit, and then subjected to real-time PCR application for screening the target virulence-associated genes. Our results showed that all of 13 isolates were positive for the virulence genes OmpA, OmpX, zpx, and Cpa. In summary, our study revealed that some of the analyzed foods and environmental samples were contaminated with pathogenic C. sakazakii with its virulence-associated markers, far above the allowable limit; and therefore, this level of contamination may pose a severe health threat for newborns, infants, and children.

Abstract Presence of extended spectrum beta-lactamases (ESBL) in bacteria is a growing health concern of global significance. The local, regional, national, and international epidemiological studies for extended spectrum beta-lactamases-producing Enterobacteriaceae and their encoding genes in foods are still incomplete. The objective of this study was to determine the occurrence of extended spectrum beta-lactamases-producing Enterobacteriaceae and the characteristics of their encoding genes from a total of 250 samples of various foods of animal-origin (100 raw chicken meat, 100 raw cow milk, and 50 raw cow milk cheese) sold in Turkey. Overall, 55 isolates were positive as extended spectrum beta-lactamases-producing Enterobacteriaceae. The most prevalent extended spectrum beta-lactamases-producing strain were identified as Escherichia coli (80%), followed by Enterobacter cloacae (9.1%), Citrobacter braakii (5.5%), Klebsiella pneumoniae (3.6%), and Citrobacter werkmanii (1.8%) by Vitek® MS. The simultaneous production of extended spectrum beta-lactamases and AmpC was detected in five isolates (9.1%) in E. coli (80%) and E. cloacae (20%). The frequency rates of blaTEM, blaCTX-M, and blaSHV were 96.4%, 53.7%, and 34.5%, respectively. The co-existence of bla -genes was observed in 82% of extended spectrum beta-lactamases producers with a distribution of blaTEM & blaCTX-M (52.7%), blaTEM & blaSHV (20%), blaTEM & blaCTX-M & blaSHV (12.7%), and blaSHV & blaCTX-M (1.8%). The most prevalent variant of blaCTX-M clusters was defined as blaCTX-M-1 (97.2%), followed by blaCTX-M-8 (2.8%). In summary, the analysed foods were found to be posing a health risk for Turkish consumers due to contamination by Enterobacteriaceae with a diversity of extended spectrum beta-lactamases encoding genes.