Unveiling Safety Concerns in Brazilian Artisanal Cheeses: A Call for Enhanced Ripening Protocols and Microbiological Assessments.

Brazilian artisanal cheeses have recently gained significant commercial prominence and consumer favor, primarily due to their distinctive sensory attributes and cultural and historical appeal. Many of these cheeses are made with raw milk and undergo a relatively short ripening period, sometimes ranging from 4 to 8 days, though it is usually shorter than the period stated by law. Moreover, there is insufficient evidence regarding the efficacy of a short ripening period in reducing certain zoonotic foodborne pathogens, such as Brucella spp., Coxiella burnetiid, and Mycobacterium bovis (as part of the Mycobacterium tuberculosis complex). Additionally, a literature analysis revealed that the usual ripening conditions of Brazilian artisanal cheeses made with raw milk may be inefficient in reducing the levels of some hazardous bacterial, including Brucella spp., Listeria monocytogenes, coagulase-positive Staphylococcus, Salmonella, and Coxiella burnetti, to the acceptable limits established by law, thus failing to ensure product safety for all cheese types. Moreover, the assessment of the microbiological safety for this type of cheese should be broader and should also consider zoonotic pathogens commonly found in bovine herds. Finally, a standardized protocol for evaluating the effectiveness of cheese ripening must be established by considering its peculiarities.

Bacteriocinogenic anti-listerial properties and safety assessment of Enterococcus faecium and Lactococcus garvieae strains isolated from Brazilian artisanal cheesemaking environment.

AIMS: This study aimed to prospect and isolate LAB from an artisanal cheese production environment, to assess their safety, and to explore their bacteriocinogenic potential against Listeria monocytogenes. METHODS AND RESULTS: Samples were collected from surfaces of an artisanal-cheese production facility and after rep-PCR and 16S rRNA sequencing analysis, selected strains were identified to be belonging to Lactococcus garvieae (1 strain) and Enterococcus faecium (14 isolates, grouped into 3 clusters) associated with different environments (worktables, cheese mold, ripening wooden shelves). All of them presented bacteriocinogenic potential against L. monocytogenes ATCC 7644 and were confirmed as safe (γ-hemolytic, not presenting antibiotic resistance, no mucus degradation properties and no proteolytic or gelatinase enzyme activity). Additionally, cell growth, acidification and bacteriocins production kinetics, bacteriocin stability in relation to different temperatures, pH and chemicals were evaluated. According to performed PCR all studied strains generated positive evidence for presence of entA and entP genes (for production of enterocins A and enterocins P, respectively). However, pediocin PA-1 associated gene were recorded only DNA from E. faecium ST02JL and Lc. garvieae ST04JL. CONCLUSIONS: It is worth considering the application of these safe LAB or their bacteriocins in situ as an alternative means of controlling L. monocytogenes in cheese production environments alone or in combination with other antimicrobials.

Revolutionizing food safety with electrochemical biosensors for rapid and portable pathogen detection.

Foodborne diseases remain a worldwide concern, despite the advances made in sanitation, pathogen surveillance and food safety management systems. The methods routinely applied for detecting pathogens in foods are time consuming, labor intensive and usually require trained and qualified individuals. The objective of this review was to highlight the use of biosensors, with a focus on the electrochemical devices, as promising alternatives for detecting foodborne pathogens. These biosensors present high speed for obtaining results, with the possibility of evaluating foods in real time, at low cost, ease of use, in addition to being compact and portable. These aspects are considered advantageous and suitable for use in food safety management systems. This work also shows some limitations for the application of biosensors, and we present perspectives with the development and use of nanomaterials.

Unveiling ochratoxin A and ochratoxigenic fungi in Brazilian artisanal Cheeses: Insights from production to consumption.

Ochratoxin A (OTA) is a toxin produced by several Aspergillus species, mainly those belonging to section Circumdati and section Nigri. The presence of OTA in cheese has been reported recently in cave cheese in Italy. As artisanal cheese production in Brazil has increased, the aim of this study was to investigate the presence of ochratoxin A and related fungi in artisanal cheese consumed in Brazil. A total of 130 samples of artisanal cheeses with natural moldy rind at different periods of maturation were collected. Of this total, 79 samples were collected from 6 producers from Canastra region in the state of Minas Gerais, since this is the largest artisanal cheese producer region; 13 samples from one producer in the Amparo region in the state of São Paulo and 36 samples from markets located in these 2 states. Aspergillus section Circumdati occurred in samples of three producers and some samples from the markets. A. section Circumdati colony counts varied from 102 to 106 CFU/g. Molecular analysis revealed Aspergillus westerdijkiae (67 %) as the most frequent species, followed by Aspergillus ostianus (22 %), and Aspergillus steynii (11 %). All of these isolates of A. section Circumdati were able to produce OTA in Yeast Extract Sucrose Agar (YESA) at 25 °C/7 days. OTA was found in 22 % of the artisanal cheese samples, ranging from 1.0 to above 1000 µg/kg, but only five samples had OTA higher than 1000 µg/kg. These findings emphasize the significance of ongoing monitoring and quality control in the artisanal cheese production process to minimize potential health risks linked to OTA contamination.

Changes in eating habits and lifestyle during the first year of the COVID-19 pandemic across metropolitan regions in Brazil and Germany: A survey-based cross-sectional study.

COVID-19 caused profound societal changes to cope rapidly with the new circumstances. The food market changed its quantity, quality, form, and frequency dynamics. Consequently, food-eating habits and lifestyles like physical exercise likely experienced changes. An online-based survey was conducted between June 2020 and January 2021 in the metropolitan regions of Rhine Ruhr Metropolis (RRM), Greater São Paulo (GSP), other metropolitan regions in São Paulo state (oMRSP), other Brazilian metropolitan regions (oBRMR), and the remaining urban areas in both countries (oUA), representing different contexts of Brazil and Germany. We assessed self-reported changes in physical activity level, diet quality, self-reported eating habits, and buying groceries during the first year of the pandemic. In Germany, indoor and outdoor activities increased for 34% of the respondents, while in Brazil, there was a decrease in physical activity for 50% of the participants. The Healthy Eating Index (HEI) scored higher among Brazilians (16.8) than Germans (15.2) on a 0-29 HEI scale. Increased awareness of healthy and sustainable eating habits was observed in GSP (0.7), oMRSP (0.63), oBRMR (0.7), and oUA (0.68) on a scale from no change (0) to change (1). In RRM, an increase in convenience foods was noticed (0.86). Participants reported discomfort with food purchasing due to hygiene measures and avoided going to the supermarket (0.7 on average in Brazil and 0.58 for females using the same 0-1 scale). Also, food supply at the grocery shops was reported to be often unavailable and in lower quantities. A real-time assessment of self-reported changes in eating habits and lifestyle during the lockdown in 2020 in different contexts is insightful for rethinking strategies to improve conditions in the post-COVID-19 era and prepare for future pandemics.

Investigating processing practices and microbiological quality of minimally processed vegetables in Brazil.

Minimally processed vegetables (MPVs) are marketed as convenient and healthy choices for consumers. However, the absence of post-commercialization treatments raises concerns about their microbiological safety. This study investigated the processing practices of 28 Brazilian MPV plants and compared the microbiological quality of these products with fresh counterparts in the city of Sao Paulo, Brazil. Through cluster analysis, the processing plants were categorized into two groups: group 1 (nineteen plants) primarily uses chemical substances in the washing step, while group 2 (nine plants) avoids chemical use but employs similar rinsing practices. Microbiological analysis of 100 samples (49 unprocessed and 51 MPVs) revealed no significant differences in microbial group counts (Enterobacteriaceae, coliforms, and E. coli) between the in natura (unprocessed) and MPV products. However, the prevalence of E. coli was higher in natura vegetables than in MPVs. The results indicated the presence of Salmonella DNA (from either dead or live cells or residual DNA) in 4 samples (3 in natura and 1 MPV) using conventional PCR, suggesting the presence of the pathogen in these samples. Listeria monocytogenes was absent, but Listeria innocua was found in two unprocessed products. The study suggests that certain MPVs have microbial loads similar to unprocessed vegetables, potentially serving as carriers for pathogen transmission. These findings emphasize the importance of understanding practices in Brazilian MPV processing plants, informing the implementation of control measures to improve MPV safety and shelf-life, thus ensuring microbiological safety.

Antimicrobial Compounds in Wine.

Ipsum vinum est potestas et possession (wine itself is power and possession). Wine is a complex system that triggers multisensory cognitive stimuli. Wine and its consumption are thoroughly intertwined with the development of human society. The beverage was appreciated in many ancient mythologies and plays an essential part in Christianity and rituals to this day. Wine has been said to enlighten and inspire artists and has even been prohibited by law and some religions, but has nevertheless played a role in human civilizations since the beginning. Winemaking is also a prospering and economically important industry and a longtime symbol of status and luxury. In winemaking, the formation of the final product is influenced by several factors that contribute to the chemical and sensory complexity often associated with quality vintages. Factors such as terroir, climatic conditions, variety of the grape, all aspects of the winemaking process to the smallest details, including metabolic processes carried out by yeast and malolactic bacteria, and the conditions for the maturation and storage of the final product, up to, and even beyond the point of deciding to open the bottle and enjoy the wine. In conjunction with the empiric and scientific process of winemaking, different molecules with antibacterial activity can be identified in wine during the production process, and several of them are clearly present in the final product. Some of these antibacterial components are phytochemicals, such as flavonoids and phenolic compounds, that may be delivered to the final product (wine) as a part of the grape, a variety of potential additive compounds, or from the oak barrels or clay amphoras used during the maturation process. Others are produced by yeasts and malolactic bacteria and play a role not only in the moderation of the fermentation process but contributing to the microbiological safety and beneficial properties spectra of the final product. Lactic acid bacteria, responsible for conducting malolactic fermentation, contribute to the final balance of the wine but are also directly involved in the production of different compounds exhibiting antibacterial activity. Some examples of these compounds include bacteriocins (antibacterial peptides), diacetyl, organic acids, reuterin, hydrogen peroxide, and carbon dioxide. Major aspects of these different beneficial metabolites are the subject of discussion in this review with the aim of highlighting their beneficial functions.

Lactic acid bacteria secreted proteins as potential Listeria monocytogenes quorum sensing inhibitors.

Listeria monocytogenes is an important human and animal pathogen able to cause an infection named listeriosis and is mainly transmitted through contaminated food. Among its virulence traits, the ability to form biofilms and to survive in harsh environments stand out and lead to the persistence of L. monocytogenes for long periods in food processing environments. Virulence and biofilm formation are phenotypes regulated by quorum sensing (QS) and, therefore, the control of L. monocytogenes through an anti-QS strategy is promising. This study aimed to identify, by in silico approaches, proteins secreted by lactic acid bacteria (LAB) potentially able to interfere with the agr QS system of L. monocytogenes. The genome mining of Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus NCFM revealed 151 predicted secreted proteins. Concomitantly, the three-dimensional (3D) structures of AgrB and AgrC proteins of L. monocytogenes were modeled and validated, and their active sites were predicted. Through protein-protein docking and molecular dynamic, Serine-type D-Ala-D-Ala carboxypeptidase and L,D-transpeptidase, potentially secreted by L. rhamnosus GG and L. acidophilus NCFM, respectively, were identified with high affinity to AgrB and AgrC proteins, respectively. By inhibiting the translocation of the cyclic autoinducer peptide (cyclic AIP) via AgrB, and its recognition in the active site of AgrC, these LAB proteins could disrupt L. monocytogenes communication by impairing the agr QS system. The application of the QS inhibitors predicted in this study can emerge as a promising strategy in controlling L. monocytogenes in food processing environment and as an adjunct to antibiotic therapy for the treatment of listeriosis.

Genomic characterization of Staphylococcus aureus from Canastra Minas Artisanal Cheeses.

Canastra Minas Artisanal Cheese is produced in the Brazilian State of Minas Gerais using raw milk, rennet, and pingo, a natural endogenous starter culture (fermented whey) collected from the previous day's production. Due to the use of raw milk, the product can carry microorganisms that may cause foodborne diseases (FBD), including Staphylococcus aureus. Genomic characterization of S. aureus is an important tool to assess diversity, virulence, antimicrobial resistance, and the potential for causing food poisoning due to enterotoxin production. This study is aimed at exploring the genomic features of S. aureus strains isolated from Canastra Minas Artisanal Cheeses. Multilocus sequence typing (MLST) classified these strains as ST1, ST5, and a new profile ST7849 (assigned to the clonal complex CC97). These strains belonged to four spa types: t008, t127, t359, and t992. We identified antimicrobial resistance genes with phenotypic correlation against methicillin (MRSA) and tetracycline. Virulome analysis revealed genes associated with iron uptake, immune evasion, and potential capacity for adherence and biofilm formation. The toxigenic potential included cyto- and exotoxins genes, and all strains presented the genes that encode for Panton-Valentine toxin and hemolysin, and two strains encoded 4 and 8 Staphylococcal enterotoxin (SE) genes. The results revealed the pathogenic potential of the evaluated S. aureus strains circulating in the Canastra region, representing a potential risk to public health. This study also provides useful information to monitor and guide the application of control measures to the artisanal dairy food production chain.

Quorum sensing interference by phenolic compounds - A matter of bacterial misunderstanding.

Over the past decade, numerous publications have emerged in the literature focusing on the inhibition of quorum sensing (QS) by plant extracts and phenolic compounds. However, there is still a scarcity of studies that delve into the specific mechanisms by which these compounds inhibit QS. Thus, our question is whether phenolic compounds can inhibit QS in a specific or indirect manner and to elucidate the underlying mechanisms involved. This study is focused on the most studied QS system, namely, autoinducer type 1 (AI-1), represented by N-acyl-homoserine lactone (AHL) signals and the AHL-mediated QS responses. Here, we analyzed the recent literature in order to understand how phenolic compounds act at the cellular level, at sub-inhibitory concentrations, and evaluated by which QS inhibition mechanisms they may act. The biotechnological application of QS inhibitors holds promising prospects for the pharmaceutical and food industries, serving as adjunct therapies and in the prevention of biofilms on various surfaces.

Minimally Processed Vegetables in Brazil: An Overview of Marketing, Processing, and Microbiological Aspects.

The global demand for minimally processed vegetables (MPVs) has grown, driven by changes in the population's lifestyle. MPVs are fresh vegetables that undergo several processing steps, resulting in ready-to-eat products, providing convenience for consumers and food companies. Among the processing steps, washing-disinfection plays an important role in reducing the microbial load and eliminating pathogens that may be present. However, poor hygiene practices can jeopardize the microbiological quality and safety of these products, thereby posing potential risks to consumer health. This study provides an overview of minimally processed vegetables (MPVs), with a specific focus on the Brazilian market. It includes information on the pricing of fresh vegetables and MPVs, as well as an examination of the various processing steps involved, and the microbiological aspects associated with MPVs. Data on the occurrence of hygiene indicators and pathogenic microorganisms in these products are presented. The focus of most studies has been on the detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes, with prevalence rates ranging from 0.7% to 100%, 0.6% to 26.7%, and 0.2% to 33.3%, respectively. Foodborne outbreaks associated with the consumption of fresh vegetables in Brazil between 2000 and 2021 were also addressed. Although there is no information about whether these vegetables were consumed as fresh vegetables or MPVs, these data highlight the need for control measures to guarantee products with quality and safety to consumers.

N-acetylcysteine (NAC) attenuates quorum sensing regulated phenotypes in Pseudomonas aeruginosa PAO1.

The expression of many virulence genes in bacteria is regulated by quorum sensing (QS), and the inhibition of this mechanism has been intensely investigated. N-acetylcysteine (NAC) has good antibacterial activity and is able to interfere with biofilm-related respiratory infections, but little is known whether this compound has an effect on bacterial QS communication. This work aimed to evaluate the potential of NAC as a QS inhibitor (QSI) in Pseudomonas aeruginosa PAO1 through in silico and in vitro analyses, as well as in combination with the antibiotic tobramycin. Initially, a molecular docking analysis was performed between the QS regulatory proteins, LasR and RhlR, of P. aeruginosa with NAC, 3-oxo-C12-HSL, C4-HSL, and furanone C30. The NAC sub-inhibitory concentration was determined by growth curves. Then, we performed in vitro tests using the QS reporter strains P. aeruginosa lasB-gfp and rhlA-gfp, as well as the expression of QS-related phenotypes. Finally, the synergistic effect of NAC with the antibiotic tobramycin was calculated by fractional inhibitory concentrations index (FICi) and investigated against bacterial growth, pigment production, and biofilm formation. In the molecular docking study, NAC bound to LasR and RhlR proteins in a similar manner to the AHL cognate, suggesting that it may be able to bind to QS receptor proteins in vivo. In the biosensor assay, the GFP signal was turned down in the presence of NAC at 1000, 500, 250, and 125 µM for lasB-gfp and rhlA-gfp (p < 0.05), suggesting a QS inhibitory effect. Pyocyanin and rhamnolipids decreased (p < 0.05) up to 34 and 37%, respectively, in the presence of NAC at 125 µM. Swarming and swimming motilities were inhibited (p < 0.05) by NAC at 250 to 10000 µM. Additionally, 2500 and 10000 µM of NAC reduced biofilm formation. NAC-tobramycin combination showed synergistic effect with FICi of 0.8, and the best combination was 2500-1.07 µM, inhibiting biofilm formation up to 60%, besides reducing pyocyanin and pyoverdine production. Confocal microscopy images revealed a stronger, dense, and compact biofilm of P. aeruginosa PAO1 control, while the biofilm treated with NAC-tobramycin became thinner and more dispersed. Overall, NAC at low concentrations showed promising anti-QS properties against P. aeruginosa PAO1, adding to its already known effect as an antibacterial and antibiofilm agent.

High Level of Interaction between Phages and Bacteria in an Artisanal Raw Milk Cheese Microbial Community.

Microbial starter cultures are used in the production of many cheeses around the world, such as Parmigiano-Reggiano, in Italy, Époisses, in France, and Canastra, in Brazil, providing many of the unique features of these cheeses. Bacteriophages (phages) are ubiquitous and well known to modulate the structure of bacterial communities, and recent data indicate that cheeses contain a high abundance of naturally occurring phages. Here, we analyze the viral and bacterial metagenomes of Canastra cheese: a traditional artisanal Brazilian cheese produced using an endogenous starter culture and raw milk. Over 1,200 viral operational taxonomic units were recovered using both isolated viral-like particles and complete metagenomic DNA. Common viral families identified included Siphoviridae and Myoviridae, with 40% of putative phage genomes unidentified at the family level of classification. We observed very high phage diversity, which varied greatly across different cheese producers, with 28% of phage genomes detected in only one producer. Several metagenome-assembled genomes were recovered for lactic acid-producing bacteria, as well as nonstarter bacterial species, and we identified several phage-bacterium interactions, at the strain level of resolution, varying across distinct cheese producers. We postulate that at least one bacterial strain detected could be endogenous and unique to the Canastra cheese-producing region in Brazil and that its growth seems to be modulated by autochthonous phages present in this artisanal production system. This phage-host relationship is likely to influence the fermentation dynamics and ultimately the sensorial profile of these cheeses, with implications for other similar cheese production systems around the world. IMPORTANCE Our work demonstrated a dynamic yet stable microbial ecosystem during cheese production using an endogenous starter culture. This was observed across several distinct producers and was marked by genomic evidence of continued phage-bacterium interactions, such as the presence of bacterial defense mechanisms. Furthermore, we provide evidence of unique microbial signatures for each individual cheese producer studied in the region, a fact that may have profound consequences on product traceability. This was the first effort to describe and understand the bacteriophage composition and ecological dynamics within the Brazilian Canastra cheese production system. The study of this prototypical backslopping production system provides a solid background for further mechanistic studies of the production of many cheeses around the world.

N-acetylcysteine (NAC) attenuates quorum sensing regulated phenotypes in Pseudomonas aeruginosa PAO1

The expression of many virulence genes in bacteria is regulated by quorum sensing (QS), and the inhibition of this mechanism has been intensely investigated. N-acetylcysteine (NAC) has good antibacterial activity and is able to interfere with biofilm-related respiratory infections, but little is known whether this compound has an effect on bacterial QS communication. This work aimed to evaluate the potential of NAC as a QS inhibitor (QSI) in Pseudomonas aeruginosa PAO1 through in silico and in vitro analyses, as well as in combination with the antibiotic tobramycin. Initially, a molecular docking analysis was performed between the QS regulatory proteins, LasR and RhlR, of P. aeruginosa with NAC, 3-oxo-C12-HSL, C4-HSL, and furanone C30. The NAC sub-inhibitory concentration was determined by growth curves. Then, we performed in vitro tests using the QS reporter strains P. aeruginosa lasB-gfp and rhlA-gfp, as well as the expression of QS-related phenotypes. Finally, the synergistic effect of NAC with the antibiotic tobramycin was calculated by fractional inhibitory concentrations index (FICi) and investigated against bacterial growth, pigment production, and biofilm formation. In the molecular docking study, NAC bound to LasR and RhlR proteins in a similar manner to the AHL cognate, suggesting that it may be able to bind to QS receptor proteins in vivo. In the biosensor assay, the GFP signal was turned down in the presence of NAC at 1000, 500, 250, and 125 μM for lasB-gfp and rhlA-gfp (p < 0.05), suggesting a QS inhibitory effect. Pyocyanin and rhamnolipids decreased (p < 0.05) up to 34 and 37%, respectively, in the presence of NAC at 125 μM. Swarming and swimming motilities were inhibited (p < 0.05) by NAC at 250 to 10000 μM. Additionally, 2500 and 10000 μM of NAC reduced biofilm formation. NAC-tobramycin combination showed synergistic effect with FICi of 0.8, and the best combination was 2500–1.07 μM, inhibiting biofilm formation up to 60%, besides reducing pyocyanin and pyoverdine production. Confocal microscopy images revealed a stronger, dense, and compact biofilm of P. aeruginosa PAO1 control, while the biofilm treated with NAC-tobramycin became thinner and more dispersed. Overall, NAC at low concentrations showed promising anti-QS properties against P. aeruginosa PAO1, adding to its already known effect as an antibacterial and antibiofilm agent.

Proposta de Pesquisa Científica para Avaliação da Segurança Microbiológica de Queijos Artesanais fabricados com leite cru e maturados por período inferior a 60 dias produzidos no estado de São Paulo / Scientific Research Proposal for the Microbiological Safety Evaluation of Raw Milk Artisanal Cheeses Ripened for Less than 60 Days in the State of São Paulo

Introdução: No Brasil, a legislação exige maturação mínima de 60 dias para queijos produzidos com leite cru, salvo se houver comprovação científica de que a redução do prazo não compromete a segurança do produto, como ocorreu com alguns queijos artesanais tradicionais de diferentes regiões do estado de Minas Gerais. No estado de São Paulo, onde os produtores seguem formulações e processos específicos individuais, a falta de um padrão de pesquisa validado dificulta a avaliação da segurança microbiológica deste tipo de queijo, pois essas pesquisas são dispendiosas e não há garantia de que os parâmetros estabelecidos pelos pesquisadores seriam aceitos pelos serviços de inspeção. Objetivo: Para abordar este desafio, esta proposta visa estabelecer um modelo de pesquisa científica padronizado para avaliar a segurança de queijos de leite cru maturados por menos de 60 dias. Métodos: Com base na proposta do DIPOA para classificação de Risco de Estabelecimentos, foi criado um checklist para classificação dos estabelecimentos/produtos, considerando risco do produto, volume processado e características de desempenho do estabelecimento em relação às Boas Práticas de Fabricação.  Resultados: A pontuação obtida pela aplicação do checklist na produção do queijo em questão determina o risco do produto como baixo, médio, alto ou muito alto e auxilia no estabelecimento do número mínimo de amostras a serem avaliadas mensalmente. Discussão: Espera-se que essa pesquisa auxilie produtores e órgãos de registro, fiscalização e inspeção no processo de legalização dos queijos artesanais de leite cru, possibilitando o desenvolvimento rural e a fixação de mão de obra no campo. (AU)

Introduction: In Brazil, the legislation requires a minimum ripening period of 60 days for raw milk cheeses, unless scientific evidence shows that the reduction in the ripening time does not compromise food safety, as seen with traditional artisanal cheeses from the state of Minas Gerais. In the state of Sao Paulo, where many producers have developed their own specific formulations and processes, the lack of a standardized validated research protocol for these types of cheese hinders their microbiological safety evaluation, since this type of research is costly and there are no guarantees that the parameters stablished by the researchers would be accepted by the inspection services. Objectives: To address this challenge, this proposal aims to stablish a standardized scientific research protocol for assessing the safety of raw milk cheeses ripened for less than 60 days. Methods: Based on the DIPOA (Department of Inspection of Products of Animal Origin) risk classification model for establishments, a checklist was created to categorize establishments/products, taking into consideration the product's risk, processed volume, and the establishment's performance assessed on the basis of good manufacturing practices. Results: The score obtained through the checklist application determines the final risk level of the product, categorized as low, medium, high, or very high, and helps determining the minimum number of samples to be evaluated monthly. Discussion: This research protocol may help producers, regulatory, inspection, and registration bodies in the legalization process of raw milk artisanal cheeses, enabling rural development and fixation of workforce in rural settings. (AU)

Labeling of ready-to-eat minimally processed vegetables sold in Brazil / Rotulagem de vegetais minimamente processados prontos para consumo comercializados no Brasil

Introduction: Ready-to-eat minimally processed vegetables (RTE-MPV) are vegetables subjected to several steps that modify their natural structure, while maintaining the same freshness and nutritional quality as the fresh produce. Since these products are sold in packages, they must be labeled, even though nutritional labeling is optional. Objective: The goal of this study was to assess the labeling aspects of several brands of RTE-MPV sold in Brazil, determining whether manufacturers adhered to the different types of food labeling legislation. Method: Photographic records of RTE-MPV packages were obtained in different regions of Brazil between October 2020 and August 2021, and labels were analyzed using a checklist that was prepared according to the different types of Brazilian food labeling legislation in force at the time of the study: RDC nº 259/2002, RDC nº 359/2003, RDC nº 360/2003 and Law nº 10,674/2003. Results: The labels of 288 RTE-MPV packages, belonging to 39 brands, were analyzed. Among these, 31 brands showed at least one aspect that was not in accordance with the legislation, such as the lack of information about place of origin, and the presence or absence of gluten. Although optional, most brands (38) adopted nutritional labeling, but the information was incomplete in ten of them. Conclusion: These data indicate that there are flaws in the labeling of RTE-MPV in Brazil, emphasizing the need for manufacturers to comply with the legislation. Moreover, the optional adoption of nutritional labeling by most brands is significantly important for consumers to have additional information about what they consume. (AU)

Introdução: Os vegetais minimamente processados (VMP) são submetidos a etapas que modificam sua estrutura natural, mantendo o frescor e qualidade nutricional dos produtos frescos. Por serem comercializados embalados, esses produtos devem ser rotulados, embora a rotulagem nutricional seja opcional. Objetivo: Este estudo analisou a rotulagem de diferentes marcas de VMP comercializados no Brasil, a fim de determinar a aderência dos produtores às legislações relativas à rotulagem de alimentos. Método: Foram obtidos registros fotográficos de embalagens VMP comercializados em diferentes regiões do Brasil entre outubro de 2020 e agosto de 2021, e os rótulos foram analisados por meio de um checklist elaborado com base nas legislações brasileiras de rotulagem de alimentos vigentes no período em que o estudo foi realizado: RDC nº 259/2002, RDC nº 359/2003, RDC nº 360/2003 e Lei nº 10.674/2003. Resultados: Foram analisados os rótulos de 288 embalagens de VMP, pertencentes a 39 marcas. Dentre essas, 31 marcas apresentaram pelo menos um item que não estava de acordo com as legislações de rotulagem vigentes, como falta de informação sobre o local de origem e a presença ou ausência de glúten. Apesar de opcional, a maioria das marcas (38) adotou a rotulagem nutricional, mas em dez delas as informações estavam incompletas. Conclusão: Esses dados indicam falhas na rotulagem de VMP no Brasil, enfatizando a necessidade das empresas de cumprirem essas regulamentações. Além disso, a adoção opcional da rotulagem nutricional pela maioria das marcas tem grande importância, pois fornece informações adicionais aos consumidores sobre os produtos que consomem. (AU)

Food Safety Issues Related to Eating In and Eating Out.

Because of growing urbanization and lack of time to prepare meals at home, eating out or getting food delivered have become common trends for many people. The consumption of food from unknown sources may impose an increased chance of contamination with microbiological hazards, especially if sanitary conditions are not met. We evaluated data from health surveillance agencies and scientific articles on foodborne diseases (FBD) reported internationally according to the exposure sites. We observed that the data are influenced by cultural, political, and socioeconomic differences. For instance, in New Zealand, Australia, United States, Denmark and India, the occurrence of FBD outbreaks was greater from foods prepared in commercial establishments and street vendors than from households. Conversely, in China, countries of the European Union and Brazil, the results are the opposite. Additionally, the pandemic imposed new eating behavior patterns, increasing delivery services and foods prepared in so-called "Dark Kitchens". The underreporting and heterogeneity of data among countries prevented a precise conclusion to the question of whether homemade foods are inherently safer than foods prepared out. Nevertheless, a lower level of development in a country influences its sanitation conditions, as well as the number of street food vendors, the search for cheaper foods, and insufficient knowledge of the population on good hygiene practices, which can all increase the chances of FBD cases.

Antimicrobial and Antioxidant Activity of Apricot (Mimusopsis comersonii) Phenolic-Rich Extract and Its Application as an Edible Coating for Fresh-Cut Vegetable Preservation.

Edible coatings have several advantages in preserving foods, such as avoiding water loss, controlling microbial growth, and reducing the need for preservatives added directly to the product. Antimicrobial action can be obtained by adding antimicrobial substances including phenolic compounds commonly found in plant extracts. This study evaluated the phenolic compounds content, antioxidant and antimicrobial activity of pulp, and seed extracts of Mimusopsis comersonii (popularly known in Brazil as abrico), besides the phenolic compounds were identified and quantified in the pulp extract. Edible coatings were incorporated with pulp extract in order to evaluate the preservation of minimally processed apples and baroa potatoes against foodborne bacteria, and enzymatic browning was also determined. Myricetin-3-glucoside, quercetin-3-glucoside, and kaempferol-3-glucoside were identified as major flavonoids in the apricot pulp extract. The seed and pulp extracts inhibited all tested microorganisms, especially Staphylococcus aureus and Salmonella Typhimurium. Edible coatings added with 9% of phenolic extract showed in vitro antimicrobial activity, in addition to being effective in preventing enzymatic browning in minimally processed apples and baroa potatoes for up to 15 days of storage. They were also effective in reducing up to 2 log CFU/g of aerobic mesophiles after 15 days of storage for apples, even though no microbial inhibition in baroa potatoes was observed under the same conditions. The addition of pulp phenolic extract in edible coatings proved to be an alternative in the preservation of apples and in the antibrowning activity of minimally processed baroa potatoes.