A presente dissertação, apresentada em dois estudos, buscou verificar as condições de boas práticas de higiene e manipulação (BPHM) e de infraestrutura (IE) de oito restaurantes/lanchonetes e de sete pontos de comércio ambulante localizados na Cidade Universitária Armando de Salles Oliveira (CUASO-USP) e a qualidade higiênico-sanitária de 45 amostras de alimentos prontos para o consumo colhidas nos mesmos. O primeiro estudo traz uma abordagem exploratória e qualitativa através da aplicação de listas de verificação; o segundo analisa laboratorialmente aspectos higiênico-sanitários dos alimentos comercializados pelos estabelecimentos alvo do estudo anterior. Concluiu-se que 1) os estabelecimentos de comércio alimentício da CUASO-USP apresentaram índices regulares de cumprimento de BPHM e adequação de IE, sendo que o eixo de higiene e manipulação de alimentos mostrou-se em melhor situação quando comparado ao de infraestrutura; 2) os ambulantes analisados apresentaram melhores resultados no cumprimento das normas de BPHM e IE e na avaliação das condições higiênico-sanitárias quando comparados aos estabelecimentos fixos de comércio alimentício. Constatou-se que é possível a prática do comércio de alimentos de rua com qualidade higiênico-sanitária, sem caracterizar uma ameaça à saúde publica, desde que o empreendedor conheça e aplique os procedimentos necessários e críticos à obtenção da garantia dos produtos comercializados, assumindo responsabilidade social ao realizar o seu modo de produção mercantil simples, porém comprometido moralmente com a sociedade. / This investigate, presented in two studies, examined the good hygiene practices and handling (GHPH) and the infrastructure (IS) conditions from eight restaurants and seven street vendors located in University City Armando de Salles Oliveira (UCASO-USP) and the sanitary conditions of 45 samples of ready to eat food harvested in them. The first study provides an exploratory and qualitative approach through the application of checklists; the second laboratory analyzes the hygienic and sanitary aspects of the food sold by the previously discussed establishments. We concluded that 1) the UCASO-USP food trade establishments showed regular compliance rates of GHPH and appropriateness of IS, and the axis of hygiene and food handling proved to be better off compared to the infrastructure; 2) the street vendors analyzed showed better results in meeting the GHPH and IS standards and in the evaluation of sanitary conditions when compared to fixed establishments of food trade. It was found that it is possible to practice the street foods trade in sanitary conditions, without characterizing a public health threat, provided that the enterprising know and apply the necessary and critical procedures to obtaining the products marketed assurance, with social responsibility while make their commodity production simple mode.

Orientador: Marcelo Cristianini / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-27T18:44:38Z (GMT). No. of bitstreams: 1 Oliveira_ThalesLeandroCoutinhode_D.pdf: 2386676 bytes, checksum: bc4655fa6960e3332639a4ca75c75545 (MD5) Previous issue date: 2015 / Resumo: A redução do cloreto de sódio nas formulações de alimentos processados representa um grande desafio para indústria moderna, tendo em vista a relação estabelecida entre o consumo elevado de sódio e a ocorrência da hipertensão arterial, principal fator de risco associado à ocorrência de distúrbios do cérebro e cardiovasculares. Em produtos cárneos, além de atuar sobre aspectos sensoriais e de textura, o sal desempenha papel chave sobre sua estabilidade microbiológica. Uma simples redução pode resultar em produtos instáveis durante sua vida de prateleira. Logo, para que estratégias de redução de sal sejam implementadas de maneira segura, faz-se necessário a aplicação de tecnologias ou agentes de preservação complementares. Tecnologias emergentes não-térmicas de inativação microbiana como o processamento à Alta Pressão (HPP - High Pressure Processing) surgem como opções factíveis para tal propósito, eliminando a dependência de conservantes químicos em face à redução de sal. A adição de agentes antimicrobianos naturais como barreiras adicionais à HPP (Tecnologia de Múltiplas Barreiras), visando potencializar os efeitos conservadores, têm sido apontada como uma promissora tendência na literatura moderna. Diante deste exposto, após determinação da formulação de trabalho com teor reduzido de sódio, a pesquisa objetivou ajustar as variáveis de processo "carga de pressão" e "tempo de residência" (utilizando baixas temperaturas de 25°C) para o processamento de embutido de peito peru fatiado embalado a vácuo (tipo "Blanquet"). Foram avaliadas respostas de inativação microbiana pós-processo (reduções logarítmicas) para populações inoculadas do patógeno Listeria monocytogenes e bactérias ácido láticas deterioradoras incluindo as espécies Leuconostoc mesenteroides e Lactobacillus sakei; também foram avaliados os atributos de qualidade incluindo cor, atividade de água, pH, sinerese, textura e oxidação lipídica. Em adição, visando à utilização de tecnologia de múltiplas barreiras para conservação, foi estudado o efeito da adição do componente bioativo carvacrol em concentrações de 200ppm, na formulação do produto. Empregando estratégia de simples redução, sem a adição sais substitutos, após avaliação de aspectos físico-químicos, sensoriais e microbiológicos, concluiu-se que níveis de 30% de redução de sal (cerca de 25% menos sódio) sobre o controle formulado com 20g/kg, são viáveis; entretanto, foi evidenciada redução da estabilidade microbiológica. Após avaliação de diferentes faixas de pressão e tempo de processo, seguindo os critérios exigidos de desempenho para inativação pós-processo de Listeria monocytogenes (4-5 reduções logarítmicas), um valor de 600MPa/180 segundos a 25°C se apresentou como um tratamento adequado para o produto com baixo teor de sódio estudado, promovendo reduções logarítmicas eficazes contra o patógeno-alvo estudado e a microbiota deteriorante. Alterações instrumentais (p<0.05) em alguns dos atributos de qualidade avaliados puderam ser destacadas principalmente sinerese, oxidação lipídica e textura; porém são necessários estudos mais amplos de análise sensorial. Adicionalmente, dados apresentados demonstraram que os consumidores não foram capazes de diferir o produto low-sodium processado por alta pressão isostática (600MPa/180sec/25°C) de um não pressurizado; de fato, apesar dos efeitos instrumentais significativos, estas alterações parecem não ser identificadas pelos consumidores. A adição de uma barreira natural complementar, o carvacrol, em níveis sensorialmente aceitáveis, foi capaz de potencializar os efeitos de inativação pós-processo (reduções logarítimicas) dos grupos microbianos alvo; efeitos benéficos combinados ao longo da estocagem refrigerada do produto também foram evidenciados por meio de reduzidas taxas de crescimento e fase lag aumentada, maximizando assim o potencial de conservação. A utilização combinada de HPP e carvacrol representou uma alternativa promissora contra fenômenos de injúrias sub-letais e recuperação celular. Sugere-se que a intensidade do processo HPP (em termos de carga de pressão, tempo de processo e temperatura) necessária para inativar microrganismos em níveis requeridos, possa ser reduzida na presença de barreiras antimicrobianas adicionais, garantindo a qualidade total do produto processado em condições mais amenas. Em adição, uma série de vantagens industriais podem ser destacadas tais como a redução de custos para a instalação de equipamento inicial e de manutenção (equipamentos operando em cargas mais baixas); e maximização da produção de processamento por ciclos efetivamente encurtados (ciclos de produtividade mais elevados por hora) / Abstract: The reduction of sodium chloride in processed food formulations represents a great challenge for modern industry, considering the relationship established between high sodium intake and the occurrence of arterial hypertension, the major risk factor associated with the occurrence of cardiovascular diseases and strokes. In meat products, in addition to acting on sensory quality and texture, salt plays a key role on the microbiological stability. A simple reduction can result in unstable products during their shelf-life. Therefore, for the success of salt reduction strategies implemented safely, it is necessary the application of complementary technologies or alternative preservatives. Non-thermal emerging technologies for microbial inactivation, such as High Pressure Processing (HPP), appear as feasible alternative for this purpose, eliminating the dependence of chemical preservatives considering the reduction of salt barrier. The addition of natural antimicrobial agents as additional hurdles (Multi Hurdle Technology) to the HPP, aiming to enhance the preservative effects, has been identified as a promising trend in modern literature. Given this exposed, after determination of low-sodium working formulation, this research aimed adjust the processing variables "pressure load" and "dwell time" (using low-processing temperatures of 25°C) for sliced vacuum-packaged ready-to-eat turkey breast. Post-processing microbial inactivation responses were evaluated (by logarithmic reductions) against inoculated populations of the target pathogen Listeria monocytogenes and spoilage lactic acid bacteria including Leuconostoc mesenteroides and Lactobacillus sakei; were also evaluated quality-attributes including color, water activity, pH, syneresis, texture and lipid oxidation. In addition, in order to use multiple barriers for preservation technology, was studied the effect of adding bioactive component, carvacrol at concentrations of 200ppm in the product formulation. Employing a simple reduction strategy, without adding salt substitutes, after evaluation of physicochemical aspects, sensory and microbiological, it was concluded that levels of 30% NaCl reduction (aproximatelly 25% less sodium) on the control made with 20g/kg, are viable; however, problems with microbiological stability were evidenced. After evaluation of different pressure ranges and processing time, following the performance criteria required for post-inactivation process Listeria monocytogenes (4-5 log reduction), a value of 600MPa/180 seconds at 25°C was presented with an adequate treatment for studied product with low sodium content, promoting effective logarithmic reductions against the studied target pathogen and spoilage LAB. Instrumental changes (p<0.05) in some of the evaluated quality attributes could be revealed mainly in syneresis, lipid oxidation and texture; however, a sensory confirmation needs to be established. Additionally, data presented showed that consumers have not been able to differ low-sodium products processed by HPP (600MPa/180sec/25°C) and unpressurised; in fact, despite significant instrumental effects, these changes seems to be not identified by consumers. The addition of natural barrier, such as carvacrol at acceptable sensory levels (200ppm), was able to potentiate the effects of post-process inactivation (logarithmic reductions) of the target microbial groups; combined benefit effects along the refrigerated product storage were also evidenced by reduced growth rates and increased lag phases, thus maximizing their potential for conservation. The combination of HPP and carvacrol represented a promising weapon against phenomena of sub-lethal injuries and cell recovery. It is suggested that the intensity of the HPP process (in terms of pressure load, process time and temperature) required to inactivate microorganisms in required levels, may be reduced in the presence of additional microbial barriers, ensuring the overall quality of the processed product under mild processing conditions. In addition, a number of industrial advantages can be highlighted such as reduced costs for the initial installation and maintenance equipment (low required pressure loads) and maximization of production by effectively processing cycles shortened (higher productivity cycles per hour) / Doutorado / Tecnologia de Alimentos / Doutor em Tecnologia de Alimentos

In many countries, the authorities face extreme difficulties in monitoring and ensuring that food sold on the street is safe, that is, fit for human consumption. This is particularly the case in urban areas, where people buy food on the street because it is readily available and relatively inexpensive. The objective of this study was to determine the food safety knowledge and microbial hazard awareness of street food consumers, and to assess the bacteriological quality of selected ready-to-eat foods sold by street vendors in the Johannesburg municipality. A cross-sectional survey study was conducted and a total of 402 respondents who buy and consume street-vended foods were randomly selected at various street food vending locations. A total of 315 various street-vended samples were purchased from randomly selected street food vendors at different vending locations in Johannesburg metropolis, in order to investigate the bacteriological quality of street-vended foods. Results of the bacteriological analysis revealed that total aerobic counts ranged from 0.3*102 - 0.4*105 cfu/g in cereals and grain-based foods; 0.4*102 - 0.5*105 cfu/g in meat-, dairy- and fish-based foods and 0.7*102 - 0.9*104 cfu/g in fruit- and vegetable-based foods. None of the food samples tested positive for Salmonella spp and Staphylococcus aureus. Results of the survey showed that the majority of respondents were black males younger than 35 years. Individuals of different gender, race, level of education and monthly income groups significantly (p<0.05) differed in their responses regarding the frequency of purchasing and confidence in the safety of street-vended food. Better taste followed closely by affordability and accessibility were the most cited reasons for purchasing street-vended food / Life and Consumer Sciences / M. Sc. (Life Sciences)

Background and motivation Informal food vending has many benefits that range from food security for the urban poor, to contribution to local economies. Yet, the street food sector is not recognized and is treated as a major public health risk. This is based partly on perceptions of local authorities that street food vendors are undesirable, or are a temporary phenomenon that will disappear in due course as a result of development. Moreover, a number of studies have documented contamination of street foods with pathogenic microorganisms like Staphylococci aureus and enteric organisms. The former can grow and express virulence in foods such as meat and meat products post cooking. To date no participatory risk analysis methods have been applied to informal markets in Tshwane, to study the food value chain of ready-to-eat (RTE) chicken and quantify the risk of exposure to selected foodborne diseases, or to determine predictors for contamination of RTE chicken sold by informal vendors. Methodology A cross-sectional analytical study design was adopted to achieve the objectives of the present study. The study area was city of Tshwane, Gauteng Province, South Africa. The study population was informal vendors selling RTE chicken to commuters at taxi and bus stations and the sampling design involved cluster sampling. Markets were the sampling units and the informal vendors the units of concern. All clusters (n=13) that were identified were included in the study and all informal vendors located within the markets were invited to be part of the study. A total of 237 vendors agreed to be interviewed, have checklists filled in for their vending stalls, and provided a sample of RTE chicken (n= 237) for laboratory analysis. Participatory Risk Analysis, a novel approach to understanding food safety in data scarce environments, was used to collect data for analysis. Participatory research methods employed in this study included: check-lists and structured interviews with informal vendors, proportional piling and focus group discussions. Enumeration of bacterial colonies from RTE chicken samples was performed using 3MTM PetrifilmTM plates (3M, St. Paul. Mn, USA). Data for the demographic profile of vendors (n=237) was analysed using descriptive statistics to establish means and proportions. Thematic analysis combined with descriptive statistical analysis was used to establish and quantify the food value chain of informally traded RTE chicken. Stochastic risk analysis using @Risk 4.5 (Palisade Corp., Ithaca, N.Y.), was used to estimate the risk of staphylococcal food poisoning from consumption of street-vended RTE chicken. Univariable and multivariable regression models were developed using Stata 9.0 (StataCorp, College Station, Texas, USA) to determine predictors for contamination of RTE chicken with E. coli and coliform bacteria respectively. Results The majority of vendors in the markets studied were females (92%), and were between 25-50 years of age. Very few young people (< 25 years) were involved. More than half of the vendors (69.47%) ran the informal food outlet as a personal business. The minority were long term employees (15.04%) or were hired to help out for a short period of time (0.88%). A high level of literacy was observed among informal vendors. Assessment of hygiene practices showed a low level of compliance with regard to possession of a certificate of acceptance, washing of hands and utensils in the same container and controlling flies at the vending site. The environment in which informal vendors operated was not conducive to production of safe food. This means that Key 1 of the WHO five keys to safer food that requires that food production takes place in a clean environment was violated. Contamination of RTE chicken was associated with the use of water collected and supplied in containers of questionable microbiological status. This was because vendors also violated Key 5 of the WHO five keys to safer food, which requires that raw materials, including water used to prepare food should be of a high microbiological quality. However, high compliance was observed with regard to the following aspects: not wearing jewellery while preparing food, using cutlery to pick up RTE chicken, washable floors (concrete slabs and cemented floors), use of potable municipal water, preparing food in a closed structure (temporary or permanent) and locating vending stalls within 30m from the toilets. Four food value chains for RTE chicken, sold by informal vendors, were identified showing extensive cross-over from the formal to informal sector. This extensive cross-over, was corroborated by the fact that over 79.3% of the RTE chicken sold on informal markets in Tshwane metropolis, was sourced from formal markets such as supermarkets or wholesalers. The food value chain for RTE chicken was short, meaning that it involved very few stages from farm to fork. Furthermore the tendency was for the RTE chicken to be purchased, cooked and consumed the same day. Although the prevalence of contamination of RTE chicken with S. aureus was high (44%), the risk of staphylococcal food poisoning was estimated to be only 1.3% (90% CI: 0% - 2.7%) for each meal of RTE chicken consumed. The mean S. aureus counts in the RTE chicken was 103.6 (90% CI: 103.3 – 103.9). This level was lower than the 105 CFU/g needed to induce staphylococcal food poisoning. While contamination of RTE chicken with E. coli (6.32 %) was low, contamination with coliforms (23.21 %) was moderate. This suggests a moderate level of food hygiene. The fly population at the stalls where the RTE was on display, intermittent washing of hands, the location of the stall >30 m from the toilets and holding RTE chicken at <70 C were identified as important predictors of contamination with E. coli. Whereas, the use of potable toilets without hand washing facilities and poor hand washing practices were a positive predictor for contamination with coliforms. Conclusions The informal trade in food in Tshwane is dominated by women and for the majority of these vendors the informal trade in food, is the main source of employment. A risk communication strategy needs to take this into consideration. There is need for intervention to improve the hygiene in the informal markets and ensure that informal vendors observe the WHO five keys to safe food. The high level of literacy observed among informal vendors is good news, as it presents a situation of educated vendors; a fertile ground for initiatives aimed at improving their lives and the service they offer to their customers. There is neither a food policy aimed at ensuring food security for the urban poor nor evidence it is being implemented. This is supported by the high number of vendors operating in temporary structures. There is a need for the municipal authorities to intervene, to provide appropriate structures. Planning of new taxi ranks should include a proper food market, with facilities that meet the legal requirements for production of safe food. Informal vendors have demonstrated an awareness of the need to practice good food hygiene as proved by the compliance with certain hygienic food handling practices. The link between the formal and informal markets suggests that the informal market of RTE chicken is well-established with a reliable source of raw chicken. Furthermore, the strong inter-linkage between the formal and informal value chain for RTE chicken, further confirms the informal sector as a potential market for locally produced product. It also emphasizes its potential to impact on the economy of the city. In addition, this strong inter-linkage, suggests that the risks associated with the formal sector might be mirrored in the informal. Therefore, as the supermarket proliferation takes root in the rest of Africa, this food value chain and its associated risks might become the norm on the continent. The present study shows that participatory risk analysis is a good way to obtain data on informal markets. Such data can then be subjected to quantitative microbial risk analysis using sophisticated biostatistical techniques, to determine and quantify the microbial risks. The low risk of staphylococcal food poisoning proved that despite the poor working conditions under which the informal food vendors prepare food, they are able to produce food that is unlikely to result in food poisoning. However, due to the high prevalence of contamination of RTE chicken with S. aureus, there is a need for the informal vendors in these markets to be trained to improve hygienic food handling practices. On the positive side, the present study demonstrated that high prevalence of contamination does not always translate into high risk. The relatively low to moderate level of contamination with E. coli and coliforms respectively, suggests a low risk of enteric foodborne diseases from ingestion of RTE chicken. Given that some and not all hygiene practices were significantly associated with contamination of RTE chicken, monitoring authorities should emphasise identified predicators to reduce the risk of contamination and the subsequent risk of contracting foodborne diseases. The difference in hygiene scoring highlights that the one-size-fits all approach to monitoring of informal markets may not be effective. These need to be tailored to suit the individual informal markets. / Thesis (PhD)--University of Pretoria, 2015. / tm2015 / Paraclinical Sciences / PhD / Unrestricted

[ES] La seguridad alimentaria es una prioridad para la población y en la actualidad cobra mayor importancia por ciertas tendencias alimentarias como el consumo de alimentos crudos y la distribución generalizada de alimentos orgánicos, que pueden ser la causa de enfermedades transmitidas por alimentos. Para garantizar la seguridad alimentaria, la detección de estos microorganismos debe realizarse de manera rápida y eficiente. Par eso, el método de cultivo microbiológico se considera el oficial para la detección de estos patógenos. Sin embargo, adolece de importantes inconvenientes, ya que no solo requiere mucho tiempo, sino que también es laborioso y consume muchos recursos. Además, puede ser limitado con respecto a la detección de bacterias fisiológicamente alteradas y/o estresadas durante el almacenamiento y la conservación. En este trabajo se ha desarrollado un protocolo sencillo y rápido para la detección simultánea de E. coli, L. monocytogenes, S. aureus y S. enterica en alimentos, mediante la combinación de una etapa de co-cultivo en medio líquido y la detección por PCR múltiple. Se ha evaluado la eficiencia de varios medios de enriquecimiento y se seleccionó el agua de peptona tamponada como el medio óptimo para el co-cultivo de las cuatro bacterias diana. También se optimizaron las condiciones de PCR múltiple y se aplicaron tanto a co-cultivos como a muestras de alimentos inoculados artificialmente, lechuga orgánica y carne picada. Después de la optimización, la PCR múltiple desarrollada fue capaz de detectar las cuatro bacterias simultáneamente, hasta con una inoculación inicial de 10^0 UFC/mL. En presencia de ambas matrices alimentarias inoculadas, tras la etapa de co-cultivo, la PCR múltiple pudo detectar simultáneamente las 3 bacterias E. coli, S. enterica y L. monocytogenes, mientras que S. aureus se ha detectado por PCR simplex, a partir del mismo extracto de ADN del co-cultivo. Los resultados obtenidos permiten concluir que el uso de un paso de co-cultivo en Agua peptona tamponada, antes de la detección por PCR simple y múltiple, puede facilitar la detección simultánea de las cuatro bacterias potencialmente presentes en las matrices alimentarias. La presencia o ausencia de la bacteria diana en los alimentos se confirma en unas 30 horas, lo que reduce el tiempo requerido para la detección en comparación con el tiempo mínimo de 7 días por método cultural. Asimismo, permite reducir el número de medios de cultivo y reactivos, para el aislamiento e identificación de bacterias que no son detectadas por PCR y que no están presentes en las matrices alimentarias, lo que supone un importante ahorro económico. / [CA] La seguretat alimentària sempre és una prioritat per a la població i en l' actualitat cobra major importància per certes tendències alimentàries, com el consum d' aliments crus i la distribució generalitzada d' aliments orgànics, que poden ser la causa de malalties transmeses per aliments. Per garantir la seguretat alimentària, la detecció d' aquests microorganismes s' ha de realitzar de manera ràpida i eficient. Per a això, el mètode de cultiu microbiològic es considera l' oficial per a la detecció d' aquests patògens. Però, hi ha importants inconvenients, ja que no només requereix més temps, sinó que també és laboriós i consumeix molts recursos. A més, pot ser limitat pel que fa a la detecció de bacteris fisiològicament alterats i/o estressats durant l'emmagatzematge i la conservació. En aquest treball s'ha desenvolupat un protocol senzill i ràpid per a la detecció simultània d' E. coli, L. monocytogenes, S. aureus i S. enterica en aliments, mitjançant la combinació d' una etapa de co-cultiu en medi líquid i la detecció per PCR múltiple. S'ha avaluat l'eficiència de diversos mitjans d'enriquiment i s'ha seleccionat l'aigua de peptona tamponada com el medi òptim per al co-cultiu dels quatre bacteris diana. També es van optimitzar les condicions de PCR múltiple i es van aplicar tant a co-cultius com a mostres d'aliments inoculats artificialment, enciam orgànic i carn picada. Després de l'optimització, la PCR múltiple desenvolupada va ser capaç de detectar els quatre bacteris simultàniament, fins a una inoculació inicial de 10^0 UFC/mL. En presència d' ambdues matrius alimentàries inoculades, després l' etapa de co-cultiu, la PCR múltiple va poder detectar simultàniament els 3 bacteris: E. coli, S. enterica i L. monocytogenes, mentre que S. aureus s' ha detectat per PCR simple, a partir del mateix extracte d' ADN del co-cultiu. Els resultats obtinguts permeten concloure que l' ús d' un pas de co-cultiu en Aigua de peptona tamponada, abans de la detecció per PCR simple i múltiple, pot facilitar la detecció simultània dels quatre bacteris potencialment presents en les matrius alimentàries. La presència o absència del bacteri diana en els aliments es confirma en unes 30 hores, la qual cosa redueix el temps requerit per a la detecció en comparació amb el temps mínim de 7 dies per mètode cultural. Així mateix, permet reduir el nombre de mitjans de cultiu i reactius, per a l' aïllament i identificació de bacteris que no són detectats per PCR i que no estan presents en les matrius alimentàries, la qual cosa suposa un important estalvi econòmic. / [EN] Food safety is a priority for the population and is nowadays more important than ever due to certain dietary trends such as the consumption of raw foods and the widespread distribution of organic foods, which may be the cause of foodborne diseases. To ensure food safety, the detection of these microorganisms must be done quickly and efficiently. Although, the microbiological culture method is considered to be the official method for the detection of these food-borne pathogens, it suffers from significant drawbacks, such as time-consuming, laborious and expensive, in addition it may be limited regarding the detection of physiologically altered and/or stressed bacteria, during storage and preservation. In this work has been developed a simple and rapid protocol for the simultaneous detection of E. coli, L. monocytogenes, S. aureus and S. enterica in food, by combining a liquid co-culture step and detection by multiplex PCR. The efficiency of several enrichment media was evaluated and buffered peptone water was chosen as the optimal medium for the co-culture of the four target bacteria. Then, optimized multiplex PCR conditions were applied to both the co-cultures and the samples of artificially inoculated foods, organic lettuce and ground meat. After optimization, the developed multiplex PCR was able to simultaneously detect the four bacteria, up to an initial inoculation of 10^0 CFU/mL. In the presence of the two inoculated food matrices, after a co-culture step, the multiplex PCR could simultaneously detect the 3 bacteria: E. coli, S. enterica and L. monocytogenes, whereas, S. aureus has been detected by simplex PCR, from the same co-culture DNA template. The results obtained allow conclusion that the use of a co-culture step in Buffered Peptone Water, before detection by simplex and multiplex PCR, can facilitate the simultaneous detection of the four bacteria potentially present in the food matrices. The presence or the absence of the target bacteria in food is confirmed in approximately 30 hours, which reduce the time required for the detection compared to the minimum time of 7 days by cultural method. Also, it allows to reduce the number of culture media and reagents, for the isolation and identification of bacteria that are not detected by PCR and which are not initially present in the food matrices, which represents a significant economic savings. / Boukharouba, A. (2022). Isolation and Identification of Foodborne Pathogens of Special Interest in Food Safety [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/182828