Simultaneous detection of Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, and Escherichia coli O157:H7 in environmental water using PMA combined with mPCR

A contaminação de fórmulas infantis pode ocorrer quando os manipuladores cometem falhas operacionais ou falhas na adequação do ambiente de preparo. O objetivo da pesquisa foi avaliar a eficácia dos procedimentos operacionais padronizados executados na área de produção de fórmulas lácteas de um Hospital. Realizou-se uma pesquisa aplicada, descritiva e explicativa com avaliação do fluxo operacional e das condições físico funcionais, análise do comportamento dos manipuladores e pesquisas microbiológicas de Coliformes totais, Escherichia coli, Bactérias aeróbias mesófilas, Bacillus cereus, Yersinia enterocolitica, Listeria monicytogenes, Clostridium perfrigens, Clostridium Sulfito Redutor, Staphylococcus coagulase positiva, Salmonella spp., Enterococcous spp, Enterobacter sakazakii, Pseudomonas aeruginosa e fungos em superfícies de fórmulas infantis e água. Os procedimentos operacionais padronizados são executados em todas as etapas do processo, entretanto, foram observadas falhas humanas. Na avaliação físico-uncional, 92% estavam em conformidade; na pesquisa microbiológica de superfícies, 12% de resultados deram presenças para Enterococcus spp., Coliformes totais, Escherichia coli e Bactérias heterotróficas aeróbias mesófilas. Nas fórmulas infantis, 8,33% dos resultados mostraram presença para Coliformes totais, Bactérias heterotróficas aeróbias mesófilas e Fungos. Os procedimentos implantados asseguram a qualidade sanitária das fórmulas, mas se verificou a necessidade de treinamento dos manipuladores. Palavras-chave: Doenças Transmitidas por Alimentos. Fórmulas Infantis. Segurança Alimentar e Nutricional. Risco Sanitário. Abstract Infant formulas contamination may occur when handlers commit operational failures or failures in the environment adequacy. The objective of this research was evaluate the effectiveness of standard operating procedures performed in the production area of milk formulations of a Hospital. An applied, descriptive and explanatory research was conducted with evaluation of operational flow and physical functional conditions, analysis of the manipulators’s behavior and microbiological investigations of MPN of total Coliforms, Escherichia col and Enterococcus spp., Count of Bacillus cereus/g, Clostridium sulfite reducer, molds and yeasts, Staphylococcus coagulase positive, aerobic mesophilic bacteria and Cronobacter sakazakii, research of Yersinia enterocolitica, Listeria monocytogenes, Salmonella spp and Pseudomonas aeruginosa on surfaces, infant formulas and water. Standard operating procedures were performed at all the process stages, however, human failures have been observed. In the physical functional evaluation, 92% was in compliance; on the surface microbiological analysis, 12% of the results present Enterococcus spp., Total Coliforms, Escherichia coli and aerobic mesophilic bacteria. In infant formulas, 8.33% results showed presence for Total Coliforms, aerobic mesophilic bacteria and Fungi. The implanted procedures assured the formulas sanitary quality, but the necessity of the manipulators’s training was verified. Keywords: Foodborne Diseases. Infant Formulas. Food and Nutrition Security. Health Risk.

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Multiplex PCR and DNA array for the detection of Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella spp. targeting virulence-related genes

Annals of Microbiology ◽

10.1007/s13213-012-0526-4 ◽

2012 ◽

Vol 63 (2) ◽

pp. 725-731 ◽

Cited By ~ 9

Author(s):

Shree Prasad Thapa ◽

Ah Reum Han ◽

Jun Mo Cho ◽

Jang Hyun Hur

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Listeria Monocytogenes ◽

Bacillus Cereus ◽

Multiplex Pcr ◽

Escherichia Coli O157 ◽

Dna Array ◽

Salmonella Spp

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Assessment of a multiplex detection method for Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes in cow milk

Universitas Scientiarum ◽

10.11144/javeriana.sc24-1.aoam ◽

2019 ◽

Vol 24 (1) ◽

pp. 277-294

Author(s):

Rocio Esperanza Patiño-Burbano ◽

Ana Karina Carrascal ◽

Jorge Luis Parra-Arango ◽

José Luis Rodríguez-Bautista

Keyword(s):

Escherichia Coli ◽

Listeria Monocytogenes ◽

Detection Rate ◽

Pathogenic Bacteria ◽

Simultaneous Detection ◽

Cow Milk ◽

Escherichia Coli O157 ◽

Salmonella Spp ◽

E Coli ◽

Microbiological Methods

Raw cow milk is considered one of the most important vehicles for pathogenic bacteria like Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes. These three bacteria are responsible for foodborne diseases. Routine microbiological methods to detect these microorganisms in cow milk can be complicated and time consuming. The aim of this work was to evaluate a method to simultaneously detect Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes in experimentally contaminated cow milk. The assessed method combined a standard microbiological culture step, using a pre-enrichment medium that favors the growth of the three focal microorganisms: SEL broth, followed by a single PCR assay. A total of 43 interference bacterial strains were used to evaluate the method’s specificity. The detection rate for the microbiological method with standard culture media was 10 UFC/mL, and that of the PCR detection, following pre-enrichment in SEL broth, was 10 UFC/mL for S. enterica and L. monocytogenes and between 1 and 5 UFC/mL for E. coli O157:H7. The PCR method showed specificity for the reference strains. Simultaneous detection by multiple PCR using SEL broth was successful for the detection of S. enterica, E. coli O157:H7, and L. monocytogenes in samples of experimentally contaminated cow milk, featuring both a high detection rate and a high specificity. This approach promises to be a feasible routine procedure when testing milk samples in industry and public health control setups.

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Multiplex PCR for Simultaneous Detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in Meat Samples

Journal of Food Protection ◽

10.4315/0362-028x-68.3.551 ◽

2005 ◽

Vol 68 (3) ◽

pp. 551-556 ◽

Cited By ~ 95

Author(s):

SUSUMU KAWASAKI ◽

NAOKO HORIKOSHI ◽

YUKIO OKADA ◽

KAZUKO TAKESHITA ◽

TAKASHI SAMESHIMA ◽

...

Keyword(s):

Escherichia Coli ◽

Listeria Monocytogenes ◽

Multiplex Pcr ◽

Simultaneous Detection ◽

Detection Sensitivity ◽

Rapid Screening ◽

Escherichia Coli O157 ◽

Salmonella Spp ◽

E Coli ◽

Meat Samples

A multiplex PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in meat samples. DNA detection sensitivity for this method was 103 CFU/ml for each pathogen. When this protocol was used for the detection of each of the above pathogenic bacteria in spiked pork samples, 1 cell per 25 g of inoculated sample could be detected within 30 h. In the samples of naturally contaminated meat, Salmonella spp., L. monocytogenes, and E. coli O157:H7 were detected over the same time period. Excellent agreement was obtained for the results of multiplex PCR and the conventional culture method, which suggests that the multiplex PCR is a reliable and useful method for rapid screening of meat products for Salmonella spp., L. monocytogenes, and E. coli O157:H7 contamination.

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