Lactic acid fermented microalgae and cyanobacteria as a new source of lipid reducing compounds: assessment through zebrafish Nile red fat metabolism assay and untargeted metabolomics.

Obesity is one of the most important threats to human health. Besides existing pharmacological or clinical interventions, novel effective and largely available solutions are still necessary. Among diverse natural resources, microalgae are well known for their complexity in the production of novel secondary metabolites. At the same time, lactic acid bacteria (LAB) are known for their capacity to metabolize, through fermentation, different matrices, and consequently to modify or produce new compounds with potential bioactivity. This work aimed to study the production of fermented microalgae and cyanobacteria, and to analyse their extracts in the zebrafish Nile red fat metabolism assay. Three microalgal species (Chlorella vulgaris, Chlorococcum sp. and Arthrospira platensis) were fermented with seven strains of LAB from 4 species (Lacticaseibacillus rhamnosus, Lacticaseibacillus casei, Lactobacillus delbrueckii bulgaricus and Lacticaseibacillus paracasei), derived from the UPCCO - University of Parma Culture Collection, Parma, Italy). All the selected strains were able to ferment the selected species of microalgae, and the most suitable substrate for LAB growth was Arthrospira platensis. Extracts from fermented Chlorella vulgaris and Chlorococcum sp. reduced significantly the neutral lipid reservoirs, which was not observed without fermentations. The strongest lipid reducing effect was obtained with Arthrospira platensis fermented with Lactobacillus delbrueckii bulgaricus 1932. Untargeted metabolomics identified some compound families, which could be related to the observed bioactivity, namely fatty acids, fatty amides, triterpene saponins, chlorophyll derivatives and purine nucleotides. This work opens up the possibility of developing novel functional foods or food supplements based on microalgae, since lactic acid fermentation enhanced the production of bioactive compounds with lipid reducing activities.

Natural Antimicrobials: A Reservoir to Contrast Listeria monocytogenes.

Natural environments possess a reservoir of compounds exerting antimicrobial activity that are forms of defence for some organisms against others. Recently, they have become more and more attractive in the food sector due to the increasing demand for natural compounds that have the capacity to protect food from pathogenic microorganisms. Among foodborne pathogens, Listeria monocytogenes can contaminate food during production, distribution, or storage, and its presence is especially detected in fresh, raw food and ready-to-eat products. The interest in this microorganism is related to listeriosis, a severe disease with a high mortality rate that can occur after its ingestion. Starting from this premise, the present review aims to investigate plant extract and fermented plant matrices, as well as the compounds or mixtures of compounds produced during microbial fermentation processes that have anti-listeria activity.

The microbiota of Mozzarella di Bufala Campana PDO cheese: a study across the manufacturing process.

Introduction: Mozzarella di Bufala Campana PDO cheese (MBC) is a globally esteemed Italian cheese. The traditional cheesemaking process of MBC relies on natural whey starter culture, water buffalo's milk, and the local agroecosystem. Methods: In this study, the microbial ecology of intermediate samples of MBC production, coming from two dairies with slightly different cheesemaking technology (dairy M large producer, and dairy C medium-small), was investigated using 16S rRNA amplicon sequencing. This research aimed to provide insights into the dynamics of microbial consortia involved in various cheesemaking steps. Results and discussion: All samples, except for raw buffalo milk, exhibited a core microbiome predominantly composed of Streptococcus spp. and Lactobacillus spp., albeit with different ratios between the two genera across the two MBC producers. Notably, the microbiota of the brine from both dairies, analyzed using 16S amplicon sequencing for the first time, was dominated by the Lactobacillus and Streptococcus genera, while only dairy C showed the presence of minor genera such as Pediococcus and Lentilactobacillus. Intriguingly, the final mozzarella samples from both producers displayed an inversion in the dominance of Lactobacillus spp. over Streptococcus spp. in the microbiota compared to curd samples, possibly attributable to the alleviation of thermal stress following the curd stretching step. In conclusion, the different samples from the two production facilities did not exhibit significant differences in terms of the species involved in MBC cheesemaking. This finding confirms that the key role in the MBC cheesemaking process lies with a small-sized microbiome primarily composed of Streptococcus and Lactobacillus spp.

Understanding the Impact of High-Pressure Treatment on Physico-Chemical, Microstructural, and Microbiological Aspects of Pumpkin Cubes.

In this study color, texture, starch-pectin, total antioxidant capacity, microbial count, and microstructure of HPP-treated Violina pumpkin cubes were evaluated. Samples were treated at six different pressures (100 to 600 MPa-HPP100 to HPP600) for 3 min. Moisture, total soluble solids, and pH showed no significant differences between untreated (UNTR) and treated samples. Pumpkin tissue showed great structural modifications as changes in cell size and shape, cell wall damage, increased cell wall thickness, cell detachment and dehydration, and calcium ions deposition mainly from HPP300 to 600. UNTR samples showed the highest value of maximum and minimum cell elongation, perimeter segment, and a more regular cell wall thickness whereas HPP600 showed the lowest values for all these parameters. A noticeable difference was observed in HPP600 samples, with a difference in terms of color (ΔE 11.3 ± 1.9) and hardness (87.4 ± 27.8 N) compared to the UNTR ones (194.9 ± 37.9 N) whereas treatments at other pressures changed less markedly the color and texture. HPP200 could ensure a higher amount of starch and pectin availability while HPP200 and HPP400 showed the highest total antioxidants capacity. High-pressure treatment from HPP400 to 600 gave the highest destruction of microorganisms but negatively influenced the structural quality as well as texture and microstructure.

Exploring the Potential of Lactic Acid Fermentation for the Recovery of Exhausted Vanilla Beans.

The market value of vanilla is constantly growing, as it is the aroma most appreciated by consumers worldwide. The key component of the aroma of vanilla beans is vanillin, which can be directly extracted from the plant, produced by chemical synthesis, or by bioconversion of natural precursors. Due to the increasing consumers' demand for products labeled as "natural," extraction from vanilla pods results in a more valuable aroma source. Once the extraction is completed, what remains are the exhausted beans that still contain small seeds and other compounds, including varying amounts of vanillin trapped in the cellular structures of the plant. The application of fermentation of exhausted vanilla beans is proposed here as a strategy to recover "natural" vanillin and other valuable aroma compounds as a result of the metabolic conversion by lactic acid bacteria (LAB). The aim of this study was to verify the fermentability of exhausted vanilla beans by-products for their valorization, allowing the recovery of high-value molecules or new applications in food products. Design of Experiment (DoE) was used to screen a library of LAB strains to identify the best condition of fermentation in response to varying cultivation conditions. A comparison between mono and co-culture of LAB was assessed. Moreover, sensory panel tests and the evaluation of the aromatic components by Solid Phase Micro Extraction-Gas Chromatography-Mass Spectrometry analysis were carried out to better understand the modification of the aroma profile after fermentation. Fermentation with LAB changed the volatile profile and sensory characteristics of the exhausted vanilla beans and represents a promising method for the valorization of these by-products.

Influence of Processing Parameters and Natural Antimicrobial on Alicyclobacillus acidoterrestris and Clostridium pasteurianum Using Response Surface Methodology.

The food industry must ensure the stability of the products, and this is often achieved by exposing foods to heat treatments that are able to ensure the absence of pathogenic or spoilage microorganisms. These treatments are different in terms of temperature and duration and could lead to a loss in nutritional and sensory value. Moreover, some types of microorganisms manage to survive these treatments thanks to the sporification process. The addition of antimicrobials can become necessary, but at present, consumers are more inclined toward natural products, avoiding synthetic and chemical additives. Antimicrobials from plants could be a valuable option and, in this context, a patent concerning an antimicrobial extract from fermented plant substrate was recently tested against foodborne pathogens revealing high antimicrobial activity. The objective of this study was the creation of a model for the evaluation and subsequent prediction of the combined effect of different process and product variables, including antimicrobial addition, on the inhibition and reduction of spore germination of target microorganisms, Alicyclobacillus acidoterrestris and Clostridium pasteurianum, responsible for spoilage of tomato-based products.

New Insights into Food Fermentation.

Food fermentation has been used for thousands of years for food preservation [...].

Pseudomonas fluorescens and Escherichia coli in Fresh Mozzarella Cheese: Effect of Cellobiose Oxidase on Microbiological Stability during Refrigerated Shelf Life.

Background: Mozzarella cheese possesses a high moisture content (50−60%) and a relatively high pH (around 5.5) and is therefore considered a perishable food product characterized by high quality deterioration and the potential risk of microbial contamination. Moreover, it can be spoiled by Pseudomonas spp. and coliform bacteria, which may be involved in different negative phenomena, such as proteolysis, discolorations, pigmentation, and off-flavors. To prevent these, different methods were investigated. In this context, the present study aims to assess the antimicrobial effect of cellobiose oxidase on Pseudomonas fluorescens (5026) and Escherichia coli (k88, k99) in mozzarella cheese during refrigerated shelf life. Methods: microbiological challenge tests were designed by contaminating the mozzarella covering liquid containing different cellobiose oxidase concentrations with P. fluorescens (5026) and E. coli (k88, k99). The behavior of these microorganisms and the variation of hydrogen peroxide concentrations were then tested under refrigerated conditions for 20 days to simulate the mozzarella cheese shelf life. Results and Conclusions: The data obtained demonstrated the effect of cellobiose oxidase on microbial growth. In particular, E. coli (k88, k99) was inhibited over the entire shelf life, while P. fluorescens (5026) was only partially affected after a few days of refrigerated storage.

Ready to eat shelf-stable brown rice in pouches: effect of moisture content on product's quality and stability.

Despite several nutritional benefits of brown rice (BR) its consumption remains limited compared to white rice. Two of the major barriers to its consumption are long cooking time and limited shelf life. However, those two hurdles can be overcome through the development of shelf-stable BR pouches to create new ready-to-eat (RTE) products, a food category that is gaining important market shares. Nevertheless, scarce information is available on the production and shelf-life stability of ready-to-eat BR products. The first objective of this study was the determination of the optimal moisture range to fully cook BR. The second objective was to determine the effect of moisture content and storage time on two fundamental parameters for consumer's acceptance of rice: color and texture. Three RTE BR pouches with moisture contents of 54%, 57% and 60% were produced and texture and color were evaluated after 1 year of storage. Significant changes in hardness and stickiness were reported during long-term storage. Moisture content negatively affected hardness and positively affected stickiness. Furthermore, storage time and moisture showed a significant effect on rice color. The present results provide information that will be useful to design new RTE meals to promote brown rice consumption.

Antimicrobial Activity of Fermented Vegetable Byproduct Extracts for Food Applications.

To prevent foodborne diseases and extend shelf-life, antimicrobial agents may be used in food to inhibit the growth of undesired microorganisms. In addition to the prevention of foodborne diseases, another huge concern of our time is the recovery of agri-food byproducts. In compliance with these challenges, the aim of this work was to more deeply investigate the antimicrobial activity of extracts derived from fermented tomato, melon, and carrot byproducts, previously studied. All the fermented extracts had antimicrobial activity both in vitro and in foodstuff, showing even higher activity than commercial preservatives, tested for comparison against spoilage microorganisms and foodborne pathogens such as Salmonella spp., L. monocytogenes, and B. cereus. These promising results highlight an unstudied aspect for the production of innovative natural preservatives, exploitable to improve the safety and shelf-life of various categories of foodstuff.

Fermentation of Agri-Food Waste: A Promising Route for the Production of Aroma Compounds.

Food waste and byproducts are generated along the entire food processing and storage chain. The large amount of waste deriving from the whole process represents not only a great economic loss but also an important ethical and environmental issue in terms of failure to recycle potentially reusable materials. New, clear strategies are needed to limit the amount of waste produced and, at the same time, promote its enhancement for further conversion and application to different industrial fields. This review gives an overview of the biological approaches used so far to exploit agri-food wastes and byproducts. The application of solid-state fermentation by different microorganisms (fungi, yeasts, bacteria) to produce several value-added products was analyzed, focusing on the exploitation of lactic acid bacteria as workhorses for the production of flavoring compounds.

Effect of fermentation with single and co-culture of lactic acid bacteria on okara: evaluation of bioactive compounds and volatile profiles.

Okara is the main soybean by-product resulting from the processing of soy milk and tofu. Despite being a product with a lot of potential and rich in many bioactive compounds such as polyphenols, it presents an unpleasant, rancid aroma. For this reason its use in the food industry is limited. In this study, we have reported the integral use of okara in a solid state fermentation process, conducted with wild strains of lactic acid bacteria, to evaluate the effect of bacterial metabolism on the volatile and polyphenolic profiles. Strains belonging to Lactobacillus acidophilus, Lacticaseibacillus rhamnosus and Pediococcus acidilactici species were used in monoculture and, for the first time, in co-culture. The results showed an improvement in the aromatic fraction showing a decrease in hexanal, responsible for off-flavour, and an increase in ketones with fruity and buttery notes in fermented okara. Polyphenols were also affected, and, in particular, a bioconversion of glucoside isoflavones to the aglycone forms was highlighted in all fermented substrates. In addition, the appearance of both phenyllactic and p-hydroxyphenyllactic acids as well as the increase in indole-3-lactic acid was observed for the first time upon okara fermentation. Overall, the co-culture appears to be the most promising for biovalorization of okara, thereby opening the possibility of its use in the development of functional ingredients.

Microbiological Contamination of Ready-to-Eat Algae and Evaluation of Bacillus cereus Behavior by Microbiological Challenge Test.

ABSTRACT: Consumption of seaweeds (forms of algae), often categorized as a superfood, is becoming popular in western countries. Algae can be marketed fresh, but are usually sold dehydrated to ensure longer shelf life. Their consumption, often as ready-to-eat, opens up possible risks for public health because of foodborne pathogens that can contaminate the raw material during harvesting or manipulation. In this study, 14 ready-to-eat foods based on dehydrated algae, representative of the most consumed species, were considered. The microbial content, with a focus on Listeria monocytogenes and Bacillus cereus, was investigated by plate counts, and B. cereus strains were isolated and identified by 16S rRNA gene sequencing. The microbiological quality was heterogeneous among the samples and, in particular, marine bacteria, Listeria spp., B. cereus, and coliforms were detected. To contribute to related risk assessment, the ability of B. cereus to grow during refrigerated storage was evaluated, to our knowledge for the first time, by a microbiological challenge test on two ready-to-eat foods based on Undaria pinnatifida and Palmaria palmata. Despite this study demonstrating the inability of B. cereus to proliferate in seaweed-based food, its presence in dehydrated foodstuffs cannot rule out replication after rehydration before consumption, making it necessary to elucidate the possible risks for consumers.

Edible Seaweeds and Spirulina Extracts for Food Application: In Vitro and In Situ Evaluation of Antimicrobial Activity towards Foodborne Pathogenic Bacteria.

Research is more and more focused on studying and selecting food preservatives of natural origin. In this scenario, algae are an excellent source of bioactive compounds, among which are antimicrobials, whose presence is variable depending on the algal species and environmental conditions. The aim of the present study was to obtain, by a food grade approach, antimicrobial extracts from five species already approved as foods and to test their efficacy in vitro (agar well diffusion assay) and in situ (microbial challenge test) towards foodborne pathogenic bacteria. Moreover, the total phenolic compounds of the extracts were determined in order to evaluate possible correlations with the antimicrobial activity. Strains belonging to Salmonella spp., Listeria monocytogenes, Escherichia coli, Staphylococcus aureus, and Bacillus cereus were considered. Overall, the extracts showed a good antimicrobial activity in vitro towards all the tested microorganisms, especially L. monocytogenes (15 mm of inhibition diameter). The effect of inhibition was monitored during 24, 48 and 120 h showing a good persistence in time. Arthrospira platensis exerted the highest efficacy, further revealed towards L. monocytogenes on salmon tartare as bacteriostatic using 0.45% of the extract and bactericidal using 0.90%. The presence of phenolic compounds could be related to the antimicrobial activity but was not revealed as the main component of this activity. The extract with the highest phenolic content (18.79 ± 1.90 mg GAE/g) was obtained from Himanthalia elongata. The efficacy, confirmed also in a food matrix, might open perspectives for their application as food preservative.

Development of Lactic Acid-Fermented Tomato Products.

BACKGROUND: lactic acid fermentation was recently proposed to produce fruit and vegetable beverages with high nutritional value. In this study, a wide screening of strains and fermentation parameters was carried out to develop fermented tomato-based drinks containing viable cells and potentially bioactive metabolites. METHODS: six different products (three extracts, two tomato juices and one tomato puree) were used as substrate for fermentation. After preliminary testing, eight fermentation conditions for each tested product were selected. The final products were stabilized with pasteurization or refrigeration and further characterized in terms of (i) antioxidant activity and (ii) total polyphenols. RESULTS: selected strains were able to grow in almost all tomato-based products except for one extract. Antioxidant activity and total phenolic content depend on products and fermentation conditions used and, except for tomato puree, an overall increase was observed. The best nutritional profile was reached in fermented samples stored at refrigerated temperature without thermal stabilization. CONCLUSION: an integrated data vision allowed to choose, for each substrate, the best combination of strains to produce novel fermented tomato-based products with different application perspectives.

Arthrospira platensis as Natural Fermentation Booster for Milk and Soy Fermented Beverages.

Arthrospira platensis, commercially known as Spirulina, is a fresh-water cyanobacterium that has been gaining increasing attention in recent years due to its high biological and nutritional value. For this reason, it has been employed in several food applications, to obtain or enhance functional and technological properties of cheese, yogurt, bread, cookies or pasta. The aim of this work was to evaluate the potential boosting effect of two different concentrations (0.25% and 0.50% w/v) of A. platensis on the fermentation capability of several starter lactic acid bacteria (LAB) strains, 1 probiotic and 4 commercial mix culture. These strains were used to ferment three different substrates and their fermentation behaviors were evaluated by impedance analyses together with rheological and color measurements. In tryptic soy broth (TSB), the A. platensis boosting effect was significantly higher if compared to yeast extract for all the starter LAB strains except for Lb. casei, which was equally stimulated. Different results were found when the same LAB strains were cultivated in SSM. The most evident boosting effect was found for S. thermophilus and Lb. casei. LAB growth was promoted by A. platensis, confirming that it could be a useful tool in the production of novel functional fermented dairy foods. The potential boosting effect was evaluated on four commercial mix cultures used to produce milk and soy fermented beverages. It was demonstrated that the booster effect took place, but it was variable and dependent not only on the mix culture used, but also on the substrate and A. platensis concentration. Also, rheological and color modifications were found to be dependent on these factors.

Antimicrobial and Fermentation Potential of Himanthalia elongata in Food Applications.

Himanthalia elongata is a brown oceanic seaweed rich in bioactive compounds. It could play an important role in food production because of its antimicrobial and antioxidant properties. Three strains belonging to the Lactobacillus casei group (Lactobacillus casei, Lactobacillus paracasei, and Lactobacillus rhamnosus) and a Bacillus subtilis strain were used for the solid-state fermentation of commercial seaweeds, and bacterial growth was monitored using the plate count method. High-pressure processing (HPP) was also employed (6000 bar, 5 min, 5 °C) before extraction. The antimicrobial activity of the extracts was tested in terms of the main food pathogenic bacteria (Salmonella spp., Listeria monocytogenes, Escherichia coli, Staphylococcus aureus, and Bacillus cereus), and the phenolic content was estimated using the Folin-Ciocalteau method. In addition, targeted UHPLC-MS2 methods were used to unravel the profile of phlorotannins. H. elongata allowed the growth of the L. casei group strains and B. subtilis, showing the fermentability of this substrate. Significant antimicrobial activity toward L. monocytogenes was observed in the extracts obtained from unfermented samples, but neither fermentation nor HPP enhanced the natural antimicrobial activity of this seaweed species. The content in the phenolic compounds decreased because of the fermentation process, and the amount of phenolics in both the unfermented and fermented H. elongata extracts was very low. Despite phlorotannins being related to the natural antimicrobial activity of this brown seaweed, these results did not support this association. Even if fermentation and HPP were not proven to be effective tools for enhancing the useful compounds of H. elongata, the seaweed was shown to be a suitable substrate for L. casei group strains as well as for B. subtilis growth, and its extracts exhibited antimicrobial activity toward foodborne pathogens.

Heat Resistance of Listeria monocytogenes in Dairy Matrices Involved in Mozzarella di Bufala Campana PDO Cheese.

The presence of Listeria monocytogenes in Mozzarella di Bufala Campana Protected Designation of Origin cheeses may depend on curd stretching conditions and post contaminations before packaging. To avoid cross-contamination, thermal treatment of water, brines and covering liquid may become necessary. The present study aimed to improve knowledge about L. monocytogenes thermal resistance focusing on the influence of some cheese making operations, namely curd stretching and heat treatment of fluids in contact with cheese after molding, in order to improve the safety of the cheese, optimize efficacy and sustainability of the processes. Moreover, the role that cheese curd stretching plays in L. monocytogenes inactivation was discussed. The 12 tested strains showed a very heterogeneous heat resistance that ranged from 7 to less than 1 Log10 Cfu/mL reduction after 8 min at 60°C. D-values (decimal reduction times) and z-values (thermal resistance constant) calculated for the most heat resistant strain among 60 and 70°C were highly affected by the matrix and, in particular, heat resistance noticeably increased in drained cheese curd. As cheese curd stretching is not an isothermal process, to simulate the overall lethal effect of an industrial process a secondary model was built. The lethal effect of the process was estimated around 4 Log10 reductions. The data provided may be useful for fresh pasta filata cheese producers in determining appropriate processing durations and temperatures for producing safe cheeses.

Solid-State Fermentation of Arthrospira platensis to Implement New Food Products: Evaluation of Stabilization Treatments and Bacterial Growth on the Volatile Fraction.

Arthrospira platensis is a cyanobacterium widely used in food formulation and mainly consumed as a food supplement because of its high amount of proteins, vitamins and minerals. Different probiotic food supplements are present in the market, and a lactic acid fermented food product like dried spirulina could be useful not only to introduce lactic acid bacteria (LAB) with beneficial effects to the diet of consumers, but also to improve or change the aromatic profile of the substrate. Therefore, the aim of this study was the evaluation of lactic acid fermentation of A. platensis biomass, focusing on the consequent changes in the aromatic profile. For this purpose, two different stabilization treatments (UV light treatment and sterilization) were applied prior to fermentation with two LAB strains, Lacticaseibacillus casei 2240 and Lacticaseibacillus rhamnosus GG. The biomass proved to be a suitable matrix for solid-state fermentation, showing a LAB growth of more than 2 log CFU/g in 48 h. The fermentation process was also useful for off-flavor reduction. In particular, the fermentation process significantly influenced the concentration of those compounds responsible for aldehydic/ethereal, buttery/waxy (acetoin and diacetyl), alkane and fermented aromatic notes (isoamyl alcohol). The heat treatment of the matrix, in addition to guaranteed safety for consumers, led to an improved aroma after fermentation. In conclusion, a fermented spirulina powder with a different aromatic profile was obtained with the applied heat treatment. Fermentation with lactic acid bacteria can be an interesting tool to obtain cyanobacterial biomasses with more pleasant sensory properties for potential use in food formulations.

Vegetable By-Product Lacto-Fermentation as a New Source of Antimicrobial Compounds.

BACKGROUND: One of the main objectives of the food industry is the shelf life extension of food products, taking into account the safety requirements and the preference of consumers attracted by a simple and clear label. Following this direction, many researchers look to find out antimicrobials from natural sources. METHODS: Tomato, carrot, and melon by-products were used as substrates for lactic acid fermentation using seven strains belonging to the Lactobacillus genus, L. plantarum, L. casei, L. paracasei, and L. rhamnosus. The obtained fermented by-products were then extracted and the antimicrobial activity toward fourteen pathogenic strains of Listeria monocytogenes, Salmonella spp., Escherichia coli, Staphylococcus Aureus, and Bacillus cereus was tested through agar well diffusion assay. RESULTS: All the extracts obtained after fermentation had highlighted antimicrobial activity against each pathogen tested. In particular, a more effective activity was observed against Salmonella spp., L. monocytogenes, S. aureus, and B. cereus, while a lower activity was observed against E. coli. CONCLUSION: Lactic acid fermentation of vegetable by-products can be a good strategy to obtain antimicrobials useful in food biopreservation.