Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition
In order to achieve bacterial inactivation in beef burgers, one of the alternatives proposed is to add organic acids as part of their formulation. The aim of this work was to evaluate the effectiveness of propidium monoazide coupled to qPCR (PMA-qPCR) as counting methodology to assess the addition o...
| Autores principales: | , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
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Institute of Food Technologists (IFT)
2025
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| Acceso en línea: | http://hdl.handle.net/20.500.12123/22016 https://ift.onlinelibrary.wiley.com/doi/10.1111/1750-3841.70109 https://doi.org/10.1111/1750-3841.70109 |
| _version_ | 1855038502267256832 |
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| author | Rey, María De Los Ángeles Cap, Mariana Favre, Leonardo Cristian Vaudagna, Sergio Ramon Mozgovoj, Marina Valeria |
| author_browse | Cap, Mariana Favre, Leonardo Cristian Mozgovoj, Marina Valeria Rey, María De Los Ángeles Vaudagna, Sergio Ramon |
| author_facet | Rey, María De Los Ángeles Cap, Mariana Favre, Leonardo Cristian Vaudagna, Sergio Ramon Mozgovoj, Marina Valeria |
| author_sort | Rey, María De Los Ángeles |
| collection | INTA Digital |
| description | In order to achieve bacterial inactivation in beef burgers, one of the alternatives proposed is to add organic acids as part of their formulation. The aim of this work was to evaluate the effectiveness of propidium monoazide coupled to qPCR (PMA-qPCR) as counting methodology to assess the addition of lactic acid (LA), caprylic acid, and fumaric acid (FA) alone and combined as inactivation agents on Escherichia coli O157 (both non-toxigenic and Shiga toxin-producing E. coli-[STEC]-strains). The effects on physicochemical properties were also evaluated. PMA-qPCR targeting stx2 and uidA genes allows quantifying STEC and non-toxigenic E. coli O157. The most effective acid combination was LA + FA for non-toxigenic E. coli O157 with a reduction of 1.06 log CFU/g measured by PMA-qPCR. However, this treatment increased hardness and chewiness once the beef burgers were cooked. Moreover, cooking weight loss was negatively affected by the addition of all organic acids tested. PMA-qPCR methodology permits to detect and quantify viable but nonculturable E. coli O157 in the beef burgers, a common limitation encountered in the traditional plate count method. The presence of bacteria in this metabolic state could explain differences among the values obtained by both methodologies. Addition of organic acids to beef burger formulations was not as effective as expected to inactivate E. coli. |
| format | info:ar-repo/semantics/artículo |
| id | INTA22016 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Institute of Food Technologists (IFT) |
| publisherStr | Institute of Food Technologists (IFT) |
| record_format | dspace |
| spelling | INTA220162025-04-23T13:35:52Z Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition Rey, María De Los Ángeles Cap, Mariana Favre, Leonardo Cristian Vaudagna, Sergio Ramon Mozgovoj, Marina Valeria Escherichia coli Carne de Res Beef PCR Organic Acids Ácido Orgánico Escherichia coli O157 Hamburguesas Monoazido de propidio Burgers Propidium monoazide In order to achieve bacterial inactivation in beef burgers, one of the alternatives proposed is to add organic acids as part of their formulation. The aim of this work was to evaluate the effectiveness of propidium monoazide coupled to qPCR (PMA-qPCR) as counting methodology to assess the addition of lactic acid (LA), caprylic acid, and fumaric acid (FA) alone and combined as inactivation agents on Escherichia coli O157 (both non-toxigenic and Shiga toxin-producing E. coli-[STEC]-strains). The effects on physicochemical properties were also evaluated. PMA-qPCR targeting stx2 and uidA genes allows quantifying STEC and non-toxigenic E. coli O157. The most effective acid combination was LA + FA for non-toxigenic E. coli O157 with a reduction of 1.06 log CFU/g measured by PMA-qPCR. However, this treatment increased hardness and chewiness once the beef burgers were cooked. Moreover, cooking weight loss was negatively affected by the addition of all organic acids tested. PMA-qPCR methodology permits to detect and quantify viable but nonculturable E. coli O157 in the beef burgers, a common limitation encountered in the traditional plate count method. The presence of bacteria in this metabolic state could explain differences among the values obtained by both methodologies. Addition of organic acids to beef burger formulations was not as effective as expected to inactivate E. coli. Para lograr la inactivación bacteriana en hamburguesas de carne de res, una de las alternativas propuestas es agregar ácidos orgánicos como parte de su formulación. El objetivo de este trabajo fue evaluar la efectividad de la monoazida de propidio acoplada a qPCR (PMA-qPCR) como metodología de conteo para evaluar la adición de ácido láctico (LA), ácido caprílico y ácido fumárico (FA) solos y combinados como agentes de inactivación en Escherichia coli O157 (tanto cepas de E. coli-[STEC] no toxigénicas como productoras de toxina Shiga). También se evaluaron los efectos en las propiedades fisicoquímicas. La PMA-qPCR dirigida a los genes stx2 y uidA permite cuantificar STEC y E. coli O157 no toxigénica. La combinación de ácidos más efectiva fue LA + FA para E. coli O157 no toxigénica con una reducción de 1.06 log UFC/g medida por PMA-qPCR. Sin embargo, este tratamiento aumentó la dureza y la masticabilidad una vez cocinadas las hamburguesas de res. Además, la pérdida de peso por cocción se vio afectada negativamente por la adición de todos los ácidos orgánicos analizados. La metodología PMA-qPCR permite detectar y cuantificar la E. coli O157 viable, pero no cultivable, en las hamburguesas de res, una limitación común en el método tradicional de recuento en placa. La presencia de bacterias en este estado metabólico podría explicar las diferencias entre los valores obtenidos con ambas metodologías. La adición de ácidos orgánicos a las formulaciones de hamburguesas de res no fue tan eficaz como se esperaba para inactivar la E. coli. Instituto Investigación Tecnología de Alimentos (ITA) Fil: Rey, María De Los Ángeles. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Tecnología de Alimentos; Argentina. Fil: Rey, María De Los Ángeles. Instituto de Ciencia y Tecnología de los Sistemas Alimentarios Sustentables (ICyTeSAS) UEDD INTA-CONICET; Argentina Fil: Cap, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Tecnología de Alimentos; Argentina. Fil: Cap, Mariana. Instituto de Ciencia y Tecnología de los Sistemas Alimentarios Sustentables (ICyTeSAS) UEDD INTA-CONICET; Argentina. Fil: Favre, Leonardo Cristian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Tecnología de Alimentos; Argentina. Fil: Favre, Leonardo Cristian. Instituto de Ciencia y Tecnología de los Sistemas Alimentarios Sustentables (ICyTeSAS) UEDD INTA-CONICET; Argentina Fil: Vaudagna, Sergio Ramon. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Tecnología de Alimentos; Argentina. Fil: Vaudagna, Sergio Ramon. Instituto de Ciencia y Tecnología de los Sistemas Alimentarios Sustentables (ICyTeSAS) UEDD INTA-CONICET; Argentina Fil: Mozgovoj, Marina Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Investigación Tecnología de Alimentos; Argentina. Fil: Mozgovoj, Marina Valeria. Instituto de Ciencia y Tecnología de los Sistemas Alimentarios Sustentables (ICyTeSAS) UEDD INTA-CONICET; Argentina 2025-04-23T13:20:34Z 2025-04-23T13:20:34Z 2025-03 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/22016 https://ift.onlinelibrary.wiley.com/doi/10.1111/1750-3841.70109 0022-1147 (print) 1750-3841 (online) https://doi.org/10.1111/1750-3841.70109 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Institute of Food Technologists (IFT) Journal of Food Science 90 (3) : e70109. (March 2025). |
| spellingShingle | Escherichia coli Carne de Res Beef PCR Organic Acids Ácido Orgánico Escherichia coli O157 Hamburguesas Monoazido de propidio Burgers Propidium monoazide Rey, María De Los Ángeles Cap, Mariana Favre, Leonardo Cristian Vaudagna, Sergio Ramon Mozgovoj, Marina Valeria Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition |
| title | Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition |
| title_full | Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition |
| title_fullStr | Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition |
| title_full_unstemmed | Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition |
| title_short | Use of propidium monoazide coupled with real-time PCR methodology to evaluate Escherichia coli O157 lethality in beef burgers after organic acid addition |
| title_sort | use of propidium monoazide coupled with real time pcr methodology to evaluate escherichia coli o157 lethality in beef burgers after organic acid addition |
| topic | Escherichia coli Carne de Res Beef PCR Organic Acids Ácido Orgánico Escherichia coli O157 Hamburguesas Monoazido de propidio Burgers Propidium monoazide |
| url | http://hdl.handle.net/20.500.12123/22016 https://ift.onlinelibrary.wiley.com/doi/10.1111/1750-3841.70109 https://doi.org/10.1111/1750-3841.70109 |
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