High-pressure processing treatment of beef burgers: Effect on Escherichia coli O157 inactivation evaluated by plate count and PMA-qPCR.

Propidium monoazide coupled to real time PCR (PMA-qPCR) is a novel methodology proposed for the quantification of viable bacteria in food after microbial inactivation treatments. The aim of this work was to assess the effectiveness of different pressure levels on the lethality of a pool of Escherichia coli O157 strains in beef burgers by plate count and PMA-qPCR using uidA as target gene. Also, the effect on native microbiota counts, E. coli O157 counts, and physiochemical parameters of beef burgers during storage in refrigeration and frozen conditions were assessed. The treatment at 600 MPa for 5 min was the most lethal and was selected for the evaluation of bacteria behavior under storage conditions. Native microbiota and E. coli O157 were not recovered during refrigerated and frozen storage (4°C for 7 days and -18°C for 35 days). Cooking weight loss, pH, chromatic parameters, and texture were affected by HPP. PRACTICAL APPLICATION: Practical Application: PMA-qPCR can be used as an alternative to assess microbial inactivation by different high pressure processing (HPP) conditions (pressure level, holding time and temperature) more rapidly than conventional plate counts. In addition, it has the benefit of being able to quantify viable but nonculturable bacteria from contaminated beef burgers after HPP. Moreover, this novel technique generates less pathogenic residues, which minimizes workers' exposure to human biohazards.

Effects of lactic, malic and fumaric acids on Salmonella spp. counts and on chicken meat quality and sensory characteristics.

The aim of this work was to assess the effectiveness of dipping chicken breast in lactic, malic and fumaric acid 3% solutions for 15 s on Salmonella counts, as well as on chicken meat quality and sensory characteristics. All three treatments effectively reduced Salmonella counts. The values of Salmonella log reduction were 2.22, 1.55 and 1.30 log CFU/g for fumaric, malic and lactic treatments, respectively. Although fumaric acid was the most effective for reducing Salmonella counts, chicken meat quality and sensory characteristics were significantly affected, even in cooked samples. Conversely, malic and lactic acids treatments caused minimal changes in chicken meat quality and sensory characteristics compared to control samples. This study shows effective alternatives to reduce Salmonella contamination on chicken breast fillets, although further studies should be considered to improve the effects on quality and sensory attributes.

Combination of organic acids and low-dose gamma irradiation as antimicrobial treatment to inactivate Shiga toxin-producing Escherichia coli inoculated in beef trimmings: Lack of benefits in relation to single treatments.

The aim of this study was to assess the efficacy of lactic acid (LA), caprylic acid (CA), high- (HDI) and low- (LDI) dose gamma irradiation and LDI combined with LA or CA on the inactivation of a pool of Shiga toxin-producing Escherichia coli (STEC) strains inoculated on beef trimmings. The three most efficacious treatments were selected to study their effect on meat quality parameters and sensory attributes. The inoculum included five native STEC serogroups (O26, O103, O111, O145 and O157). The treatments applied were 0.5% LA, 0.04% CA, 0.5 kGy LDI, 2 kGy HDI, LDI+LA and LDI+CA. Beef trimmings were divided into two groups; one was inoculated with high (7 log CFU/g) and the other with low (1 log CFU/g) level of inoculum. Efficacy was assessed by estimating log reduction and reduction of stx- and eae-positive samples after enrichment, respectively. Results showed that treatments with organic acids alone were not effective in reducing STEC populations. For high inoculum samples, the most effective treatment was HDI followed by LDI+LA and LDI alone or combined with CA. For low inoculum samples, the most effective treatment was HDI followed by LDI alone or combined with organic acids. Concerning meat quality parameters and sensory attributes, irradiation treatments (LDI and HDI) caused minimal changes, while LDI+LA modified them significantly compared with the control. Therefore, based on our results, no benefits were observed after combining organic acids with gamma irradiation.

Inactivation of Shiga toxin-producing Escherichia coli in fresh beef by electrolytically-generated hypochlorous acid, peroxyacetic acid, lactic acid and caprylic acid.

Several studies have been conducted to verify the decontamination potential of electrolytically-generated hypochlorous acid, peroxyacetic acid, lactic acid and caprylic acid against Shiga toxin-producing Escherichia coli (STEC) in beef products. However, there is no consensus regarding their effectiveness. The aim of this study was to compare these four treatments under the same conditions and establish a ranking according to their effectiveness to inactivate STEC in fresh beef. Samples were inoculated with two levels of inoculum and rinsed for 15 s in 100 mL of antimicrobial solution treatment. Caprylic acid was the most effective treatment, followed by lactic acid and peroxyacetic acid. Electrolytically-generated hypochlorous acid had no effect. Sensory analysis showed no significant differences either in flavor or in color between samples treated with caprylic acid and reference samples. Caprylic acid appears to be an effective and viable alternative to conventional interventions frequently used for meat product decontamination.

Frecuencia, caracterización y análisis genotípico de Escherichia coli productor de toxina Shiga en frigoríficos de carne bovina de Argentina / Frequency, characterization and genotypic analysis of Shiga toxin-producing Escherichia coli in beef slaughterhouses of Argentina

The objectives of this study were: (1) to estimate STEC frequency in hide and carcass samples taken from beef slaughterhouses supplying the domestic market in Argentina, (2) to establish the pheno-genotypic characteristics of STEC and non-toxigenic Escherichia coli of serogroups O26, O45, O103, O121, O111, O145 or O157 isolated from the analyzed samples and, (3) to study their clonal relatedness. Sixty hides and 60 carcasses were analyzed. At the screening step, 48% of hide and 80% of carcass samples tested positive for the stx gene by endpoint PCR. The STEC isolation rate was 5% for hides and 8% for carcasses. The isolation rate of STEC-positive for O26, O45, O103, O111, O145 or O157 serogroups was 0% for hides and 2% for carcasses. With the purpose of studying the clonal relatedness of isolates, macrorestriction fragment analysis by pulsed-field gel electrophoresis was performed. The results indicated cross-contamination between hides and between carcasses of animals in the same lot and, that the origin of carcass contamination was their own hide, or the hides of other animals in the same lot. The high detection rate at the screening step, especially in carcasses, and the evidence of cross-contamination show the need to apply additional in-plant intervention strategies aimed at preventing carcass contamination.

Los objetivos del presente estudio fueron tres: 1) estimar la frecuencia de Escherichia coli productor de toxina Shiga (STEC) en muestras de cuero y carcasa de bovinos en frigoríficos de consumo interno de Argentina; 2) realizar la caracterización feno-genotípica de las cepas STEC y de Escherichia coli no toxigénicas pertenecientes a los serogrupos O26, O45, 0103, O121, O145 u O157 aisladas a partir de las muestras analizadas; 3) establecer la relación clonal de ese conjunto de cepas. Se analizaron 60 cueros y 60 carcasas. En la etapa de tamizaje, el gen stx se detectó en el 48% de las muestras de cuero y en el 80% de las muestras de carcasa por una PCR de punto final. La frecuencia de recuperación de cepas STEC fue del 5% en cueros y del 8% en carcasas, y la de cepas STEC positivas para los serogrupos O26, O45, O103, O121, O111, O145 u O157 fue del 0% en los cueros y del 2% en las carcasas. La relación clonal de las cepas aisladas se investigó a través de electroforesis de campo pulsado y análisis de los patrones de macrorrestricción generados. Los resultados demostraron la existencia de contaminación cruzada entre cueros y carcasas de animales pertenecientes a un mismo lote, y también que el origen de la contaminación fue el propio cuero del animal o el cuero de otros animales pertenecientes al mismo lote. Los altos porcentajes de detección en la etapa de tamizaje, especialmente en carcasas, y la evidencia de contaminación cruzada ponen de manifiesto la necesidad de evaluar la implementación de estrategias de intervención tendientes a evitar la contaminación de carcasas.

Frequency, characterization and genotypic analysis of Shiga toxin-producing Escherichia coli in beef slaughterhouses of Argentina.

The objectives of this study were: (1) to estimate STEC frequency in hide and carcass samples taken from beef slaughterhouses supplying the domestic market in Argentina, (2) to establish the pheno-genotypic characteristics of STEC and non-toxigenic Escherichia coli of serogroups O26, O45, O103, O121, O111, O145 or O157 isolated from the analyzed samples and, (3) to study their clonal relatedness. Sixty hides and 60 carcasses were analyzed. At the screening step, 48% of hide and 80% of carcass samples tested positive for the stx gene by endpoint PCR. The STEC isolation rate was 5% for hides and 8% for carcasses. The isolation rate of STEC-positive for O26, O45, O103, O111, O145 or O157 serogroups was 0% for hides and 2% for carcasses. With the purpose of studying the clonal relatedness of isolates, macrorestriction fragment analysis by pulsed-field gel electrophoresis was performed. The results indicated cross-contamination between hides and between carcasses of animals in the same lot and, that the origin of carcass contamination was their own hide, or the hides of other animals in the same lot. The high detection rate at the screening step, especially in carcasses, and the evidence of cross-contamination show the need to apply additional in-plant intervention strategies aimed at preventing carcass contamination.