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.

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.

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)

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.

Obtaining Bioproducts from the Studies of Signals and Interactions between Microalgae and Bacteria.

The applications of microalgae biomass have been widely studied worldwide. The classical processes used in outdoor cultivations of microalgae, in closed or open photobioreactors, occur in the presence of bacteria. Understanding how communication between cells occurs through quorum sensing and evaluating co-cultures allows the production of microalgae and cyanobacteria to be positively impacted by bacteria, in order to guarantee safety and profitability in the production process. In addition, the definition of the effects that occur during an interaction, promotes insights to improve the production of biomolecules, and to develop innovative products. This review presents the interactions between microalgae and bacteria, including compounds exchanges and communication, and addresses the development of new pharmaceutical, cosmetic and food bioproducts from microalgae based on these evaluations, such as prebiotics, vegan skincare products, antimicrobial compounds, and culture media with animal free protein for producing vaccines and other biopharmaceutical products. The use of microalgae as raw biomass or in biotechnological platforms is in line with the fulfillment of the 2030 Agenda related to the Sustainable Development Goals (SDGs).

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.

Furanone and phytol influence metabolic phenotypes regulated by acyl-homoserine lactone in Salmonella.

Salmonella is an important foodborne pathogen, and it is unable to produce the quorum sensing signaling molecules called acyl-homoserine lactones (AHLs). However, it synthesizes the SdiA protein, detecting AHL molecules, also known as autoinducer-1 (AI-1), in the external environment. Exogenous AHLs can regulate specific genes related to virulence and stress response in Salmonella. Thus, interfering with quorum sensing can be a strategy to reduce virulence and help elucidate the cell-to-cell communication role in the pathogens' response to extracellular signals. This study aimed to evaluate the influence of the quorum sensing inhibitors furanone and phytol on phenotypes regulated by N-dodecanoyl homoserine lactone (C12-HSL) in Salmonella enterica serovar Enteritidis. The furanone C30 at 50 nM and phytol at 2 mM canceled the alterations promoted by C12-HSL on glucose consumption and the levels of free cellular thiol in Salmonella Enteritidis PT4 578 under anaerobic conditions. In silico analysis suggests that these compounds can bind to the SdiA protein of Salmonella Enteritidis and accommodate in the AHL binding pocket. Thus, furanone C30 and phytol act as antagonists of AI-1 and are likely inhibitors of the quorum sensing mechanism mediated by AHL in Salmonella.

Exploring Phenolic Compounds as Quorum Sensing Inhibitors in Foodborne Bacteria.

The emergence of multidrug-resistant bacteria stimulates the search for new substitutes to traditional antimicrobial agents, especially molecules with antivirulence properties, such as those that interfere with quorum sensing (QS). This study aimed to evaluate the potential of phenolic compounds for QS inhibition in a QS biosensor strain (Chromobacterium violaceum) and three foodborne bacterial species (Aeromonas hydrophila, Salmonella enterica serovar Montevideo, and Serratia marcescens). Initially, an in silico molecular docking study was performed to select the compounds with the greatest potential for QS inhibition, using structural variants of the CviR QS regulator of C. violaceum as target. Curcumin, capsaicin, resveratrol, gallic acid, and phloridizin presented good affinity to at least four CviR structural variants. These phenolic compounds were tested for antimicrobial activity, inhibition of biofilm formation, and anti-QS activity. The antimicrobial activity when combined with kanamycin was also assessed. Curcumin, capsaicin, and resveratrol inhibited up to 50% of violacein production by C. violaceum. Biofilm formation was inhibited by resveratrol up to 80% in A. hydrophila, by capsaicin and curcumin up to 40% in S. Montevideo and by resveratrol and capsaicin up to 60% in S. marcescens. Curcumin completely inhibited swarming motility in S. marcescens. Additionally, curcumin and resveratrol increased the sensitivity of the tested bacteria to kanamycin. These results indicate that curcumin and resveratrol at concentrations as low as 6µM are potential quorum sensing inhibitors besides having antimicrobial properties at higher concentrations, encouraging applications in the food and pharmaceutical industries.

Plant compounds and nonsteroidal anti-inflammatory drugs interfere with quorum sensing in Chromobacterium violaceum.

Chromobacterium violaceum is a Gram-negative, saprophytic bacterium that can infect humans and its virulence may be regulated by quorum sensing via N-acyl homoserine lactones. A virtual screening study with plant compounds and nonsteroidal anti-inflammatory drugs for inhibition of C. violaceum quorum sensing system has been performed. In vitro evaluation was done to validate the in silico results. Molecular docking showed that phytol, margaric acid, palmitic acid, dipyrone, ketoprofen, and phenylbutazone bound to structures of CviR proteins of different C. violaceum strains. Phytol presented higher binding affinities than AHLs and furanones, recognized inducers, and inhibitors of quorum sensing, respectively. When tested in vitro, phytol at a non-inhibitory concentration was the most efficient tested compound to reduce phenotypes regulated by quorum sensing. The results indicate that in silico compound prospection to inhibit quorum sensing may be a good tool for finding alternative lead molecules.

Brazilian Artisanal Cheeses: Diversity, Microbiological Safety, and Challenges for the Sector.

Artisanal cheeses made with raw milk are highly appreciated products in Brazil. Most of these cheeses are produced in small facilities across different production regions in the country, some of which have been granted a protected designation of origin and are award winners. The most prominent state that manufactures these products is Minas Gerais (MG), but production is also gaining strength in other Brazilian states. The major challenge faced by artisanal cheese production is related to microbial risks associated with foodborne pathogens when the quality of the raw milk is unsatisfactory. Regulations created for the dairy industry are constantly been revised and adapted, considering the small-scale production of Brazilian artisanal cheeses, in order to guarantee safety at all steps of cheese production and commercialization. This text presents a summary of the huge diversity of artisanal cheeses produced in the country, grouped by geographical regions, and reviews the current challenges faced by producers and government considering the safety of these cheeses.

Listeria monocytogenes inhibition by lactic acid bacteria and coliforms in Brazilian fresh white cheese.

INTRODUCTION: Minas fresh cheese (MFC), a Brazilian white cheese, is one of the most popular cheeses nationwide. Studies have shown that Listeria monocytogenes occurrence in this product is generally low, while high populations of coliforms can be found. This study aimed to evaluate the influence of coliforms in the behavior of L. monocytogenes in MFC. METHODS: Pasteurized milk was inoculated with L. monocytogenes and coliforms, and the acidification was made by lactic acid or by the addition of a starter culture. The cheeses of each production were divided into 3 groups and stored at 5 ºC, 12 ºC and cycles of 5 ºC followed by 25 ºC. In predetermined days, samples were taken and L. monocytogenes, coliforms and lactic acid bacteria populations were evaluated, besides the pH, water activity (aw), titratable acidity and NaCl concentration. RESULTS: The inhibition of L. monocytogenes in the presence of coliforms was observed (p < 0.05), except for those samples prepared with lactic acid and stored at temperature cycles. The values of pH and aw were not sufficiently low to cause inhibition; however, titratable acidity was higher in cheeses containing coliforms. In vitro tests containing lactic acid and L. monocytogenes showed that the bacterium is sensitive to concentration of lactic acid ≥ 0.3%, indicating that lactic acid produced by coliforms strongly influences the population of L. monocytogenes. CONCLUSIONS: Thus, it can be concluded that coliforms negatively impact populations of L. monocytogenes in MFC. We strongly recommend that producers of MFC adopt good hygiene practices to not only avoid contamination with L. monocytogenes, but also coliforms.

Phenolic extract of Eugenia uniflora L. and furanone reduce biofilm formation by Serratia liquefaciens and increase its susceptibility to antimicrobials.

Serratia liquefaciens is a spoilage microorganism of relevance in the dairy industry because it is psychrotrophic, able to form biofilm, and produces thermoresistant proteases and lipases. Phenolic compounds and furanones have been studied as inhibitors of biofilm formation. In this study, the potential of the pulp phenolic extract of Eugenia uniflora L. orange fruits, also called pitanga, and furanone C30 on the inhibition of biofilm formation by S. liquefaciens L53 and the susceptibility to different antimicrobials were evaluated. The pulp phenolic extract of pitanga had a high total phenolic content, being mainly composed of glycosylated quercetins and ellagitannins. Sub-inhibitory concentrations of this extract and furanone reduced biofilm formation by S. liquefaciens on polystyrene and the amount of polysaccharides, proteins and extracellular DNA in the biofilms. These biofilms were also more susceptible to kanamycin. The combinations of furanone with phenolic extract of pitanga or kanamycin showed a synergistic effect with total growth inhibition of S. liquefaciens.