No abstract

This study evaluated biofilm formation and antimicrobial susceptibility of Pseudomonas aeruginosa isolated to evaluate the microbiological quality of 80 natural mineral water samples sold in 20 L bottles. The quantity of P. aeruginosa and enterococci was assessed, including total coliforms, thermotolerant coliforms and sulfite-reducing clostridia (SRC). Biofilm production from P. aeruginosa was evaluated in brain heart infusion broth (BHI) and sterile natural mineral water at temperatures of 25 and 35°C for 24 and 48 h. Antimicrobial susceptibility tests were performed using the agar diffusion method (Kirby-Bauer). Of the 80 samples analyzed, 40 (50%) presented unsatisfactory microbiological quality according to RDC no. 275/05. Thirty-eight (47.5%) samples presented P. aeruginosa, nine (11.2%) total coliforms, four (5.0%) SRC and one (1.2%) thermotolerant coliform. Sixteen P. aeruginosa strains (51.6%) were classified as non-adherent or weakly adherent, both in BHI as in mineral water. However, five strains (16.1%) were strongly adherent in the two matrices, mainly in BHI at the temperature of 25ºC. The study also isolated resistant or intermediate resistant strains to antibiotics of aminoglycosides and/or β-lactams classes. It was concluded that P. aeruginosa isolates were able to produce biofilm in the studied matrices and presented resistance to antimicrobials. Half of the samples presented unsatisfactory microbiological quality, mostly due to P. aeruginosa contamination.

O uso de materiais de referência (MR) é uma das principais ferramentas utilizadas para garantia e controle da qualidade de laboratórios de microbiologia de alimentos. No Brasil, a RDC n.º 12/01 da Anvisa prevê como um dos parâmetros para a avaliação da qualidade de queijos a enumeração de estafilococos coagulase positiva (ECP). O grande desafio na produção de MR destinados a ensaios microbiológicos é a instabilidade natural dos micro-organismos, o que dificulta o desenvolvimento e a manutenção desses MR. O objetivo deste estudo foi produzir um MR quantitativo destinado ao ensaio de enumeração de ECP em matriz queijo. Uma amostra de queijo ultrafiltrado com contagem de ECP <10 UFC/g e número de aeróbios viáveis de 1,22 × 10³ UFC/g foi utilizada como matriz para produção do MR. A matriz foi distribuída em frascos, contaminada com a bactéria alvo em concentração específica e submetida à liofilização. Como crioprotetor, foi utilizada sacarose. O MR produzido foi considerado homogêneo e estável à temperatura < -70 ºC durante todo o período estudado (dez meses). O material apresentou estabilidade a 4, 25, 30 e 35 ºC durante quatro dias; contudo, os resultados indicam que, a 35 ºC, ocorre um decréscimo na concentração celular. A -20 ºC, o MR apresentou-se estável durante 48 dias. Conclui-se que o material apresentou todos os requisitos necessários de um MR de qualidade e poderia ser transportado aos laboratórios participantes de um ensaio de proficiência a temperaturas de até 35 ºC por até quatro dias, uma vez que os resultados indicaram a manutenção da concentração celular neste período. Esse foi o primeiro trabalho a descrever uma metodologia de produção de MR contendo ECP em matriz queijo.

No abstract

Cronobacter infections of infants are commonly regarded as due to the ingestion of contaminated feed. The aim of this study was to determine the occurrence of Cronobacter, total coliforms and Escherichia coli in different brands of natural mineral waters as sold in 20 l returnable bottles in Rio de Janeiro, Brazil. The quantification of total coliforms and E. coli was performed by Most Probable Number. The detection of Cronobacter was as according to the ISO 22964:2017 and Bacteriological Analytical Manual/FDA. Molecular characterization of Cronobacter isolates was performed by real‐time PCR and by multi‐locus sequence typing. The antibiotic susceptibility profile was determined and biofilm production was evaluated in polystyrene microplates. Total coliforms and E. coli were detected in 13 (39·4%) and 2 (6·1%) of the 33 lots analysed respectively, and were considered unsatisfactory for human consumption according to Brazilian law. One (3·0%) lot showed contamination by C. malonaticus ST440 (Cronobacter MLST Databases accession no. ID 2646). The strain was susceptible to all (n = 13) antibiotics tested and only formed a weak biofilm. Since there is a high consumption of natural mineral waters by elderly and immunosuppressed persons, epidemiological surveillance agencies should be aware of the risk that these waters may represent for these groups. Significance and Impact of the Study Cronobacter malonaticus ST440 was isolated from 20 l bottled drinking natural mineral waters sold in markets in Rio de Janeiro State, Brazil, and can be a potential threat to human health, particularly for neonates. Thirteen lots (39·4%) were unsatisfactory for human consumption due to the presence of total coliforms and/or Escherichia coli.