Microbial trace based on PCR-DGGE to evaluate the ripening stage of minas artisanal cheeses from the Canastra microregion produced by different dairies.

The Minas artisanal cheese from the Serra da Canastra (MAC-CM) microregion is a traditional product due to its production and ripening process. Artisanal chesses manufactured with raw cow's milk and endogenous dairy starters ("also known as pingo") have distinctive flavors and other sensory characteristics because of the unknown microbiota. The aim of this study was to evaluate the microbiota during 30 days of ripening, the physicochemical changes, and their relation in MACs produced in two different microregions located in the Serra da Canastra microregion through culture-dependent and culture-independent methods. The MACs were collected in the cities of Bambuí (MAC-CMB) and Tapiraí (MAC-CMT) in the Canastra microregion (n = 21). Cheeses uniqueness was demonstrated with the multivariate analysis that joined the microbiota and physicochemical characteristics, mainly to the proteolysis process, in which the MAC-CMT showed deeper proteolysis (DI -T0:14.18; T30: 13.95), while the MAC-CMB reached only a primary level (EI -T0:24.23; T30: 31.10). Abiotic factors were responsible for the differences in microbial diversity between the cheese farms. Different microbial groups: the prokaryotes, like Corynebacterium variabile, Lactococcus lactis, and Staphylococcus saprophyticus; and the eukaryotes, like Kluyveromyces lactis and Diutina catenulata dominated ripening over time. The microbial community and proteolysis were responsible for the predominance of volatile groups, with alcohols predominating in MAC-CMB and free fatty acids/acids and esters in MAC-CMT.

Emulsified and Nanoemulsified Essential Oils in the Control of Clostridium botulinum and Clostridium sporogenes in Mortadella.

Clostridium botulinum is a foodborne pathogen responsible for severe neuroparalytic disease associated with the ingestion of pre-formed toxin in food, with processed meats and canned foods being the most affected. Control of this pathogen in meat products is carried out using the preservative sodium nitrite (NaNO2), which in food, under certain conditions, such as thermal processing and storage, can form carcinogenic compounds. Therefore, the objective was to use nanoemulsified essential oils (EOs) as natural antimicrobial agents, with the aim of reducing the dose of NaNO2 applied in mortadella. The antimicrobial activity of nanoemulsions prepared with mixtures of EOs of garlic, clove, pink pepper, and black pepper was evaluated on endospores and vegetative cells of C. botulinum and Clostridium sporogenes (surrogate model) inoculated in mortadella prepared with 50 parts per million NaNO2. The effects on the technological (pH, water activity, and color) and sensory characteristics of the product were also evaluated. The combinations of EOs and their nanoemulsions showed sporicidal effects on the endospores of both tested microorganisms, with no counts observed from the 10th day of analysis. Furthermore, bacteriostatic effects on the studied microorganisms were observed. Regarding the technological and sensorial characteristics of the product, the addition of the combined EOs had a negative impact on the color of the mortadella and on the flavor/aroma. Despite the strong commercial appeal of adding natural preservatives to foods, the effects on flavor and color must be considered. Given the importance of controlling C. botulinum in this type of product, as well as the reduction in the amount of NaNO2 used, this combination of EOs represents a promising antimicrobial alternative to this preservative, encouraging further research in this direction.

Untargeted Metabolomics Approach of Cross-Adaptation in Salmonella Enterica Induced by Major Compounds of Essential Oils.

Cross-adaptation phenomena in bacterial populations, induced by sublethal doses of antibacterial solutions, are a major problem in the field of food safety. In this regard, essential oils and their major compounds appear as an effective alternative to common sanitizers in food industry environments. The present study aimed to evaluate the untargeted metabolomics perturbations of Salmonella enterica serovar Enteritidis that has been previously exposed to the sublethal doses of the major components of essential oils: cinnamaldehyde, citral, and linalool (CIN, CIT, and LIN, respectively). Cinnamaldehyde appeared to be the most efficient compound in the assays evaluating the inhibitory effects [0.06% (v/v) as MBC]. Also, preliminary tests exhibited a phenotype of adaptation in planktonic and sessile cells of S. Enteritidis when exposed to sublethal doses of linalool, resulting in tolerance to previously lethal concentrations of citral. A metabolomics approach on S. Enteritidis provided an important insight into the phenomenon of cross-adaptation induced by sublethal doses of major compounds of some essential oils. In addition, according to the results obtained, when single molecules were used, many pathways may be involved in bacterial tolerance, which could be different from the findings revealed in previous studies regarding the use of phytocomplex of essential oils. Orthogonal projection to latent structures (OPLS) proved to be an interesting predictive model to demonstrate the adaptation events in pathogenic bacteria because of the global engagement to prevent and control foodborne outbreaks.

Combined effects of yacon flour and probiotic yogurt on the metabolic parameters and inflammatory and insulin signaling proteins in high-fat-diet-induced obese mice.

BACKGROUND: Prebiotics and probiotics may be effective dietary components that can alter the gut microbiota of the host and, consequently, overcome imbalances associated with obesity. This work aimed to evaluate the synergistic and isolated effects and mechanisms by which probiotic yogurt containing Bifidobacterium animalis and/or Lactobacillus acidophilus and yacon flour alter metabolic parameters and inflammatory and insulin signaling proteins in diet-induced obese mice. Swiss mice were fed a high-fat diet (n = 48) or a standard diet (control; n = 6) for 56 days. The 42 mice that gained the most weight were selected and divided into seven groups that received different combinations of probiotic yogurt and yacon flour. After 30 days, biochemical parameters (blood glucose, serum total cholesterol, and triacylglycerols), crude fat excretion in feces, and periepididymal fat were assessed and an immunoblotting analysis of insulin signaling proteins and interleukin-1ß was conducted. RESULTS: The combination of yacon flour and a yogurt with two strains of probiotics exerted positive effects on the parameters evaluated, such as decreased body weight (-6.5%; P < 0.05), fasting glucose (-23.1%; P < 0.05), and triacylglycerol levels (-21.4%; P < 0.05) and decreased periepididymal fat accumulation (-44.2%; P < 0.05). There was a decrease in inflammatory markers (P < 0.001) and an improvement in insulin signaling (P < 0.001). CONCLUSIONS: The combination of a prebiotic with two strains of probiotics in a food matrix may exert a protective effect against obesity-associated inflammation, improving insulin resistance, even in the short term. © 2022 Society of Chemical Industry.

Morphological and metabolomics impact of sublethal doses of natural compounds and its nanoemulsions in Bacillus cereus.

Microbiological safety in food industry are always a concern regarding sublethal tolerance in bacteria for common and natural sanitizers. Natural bacteriocins, such as nisin (NIS), may negatively interfere in the efficiency of major compounds of essential oils against foodborne pathogenic bacteria. However, nanoemulsioned forms increase the bactericidal potential of natural compounds acting synergistically. In this study, cinnamaldehyde (CIN), citral (CIT), and linalool (LIN) were evaluated independently, associated with NIS, and in nanoemulsions (NEs) against Bacillus cereus using untargeted-metabolomics. Results revealed morphological changes in the structure of B. cereus treated with NEs of CIN and CIT, both NIS-associated. In addition, sensibility tests and UHPLC-QTOF-MS analyses indicated that NIS might react together with CIT reducing the bactericidal efficiency, while the nanoemulsion of CIT effect was enhanced by NIS in nanoemulsioned forms. This study highlights the importance of prudent administration of natural compounds as antimicrobial agents to prevent sublethal tolerance in pathogenic bacteria.

Influence of lipid nanoparticle physical state on ß-carotene stability kinetics under different environmental conditions.

Carotenoids are lipophilic compounds that provide important health-related benefits for human body functions. However, they have low water solubility and chemical stability, hence their incorporation in aqueous-based foods requires the use of emulsion-based lipid carriers. This work aimed at elucidating whether their inclusion in emulsion-based Solid Lipid Nanoparticles (SLNs) can provide a protective effect against ß-carotene degradation under different environmental conditions in comparison to liquid lipid nanoemulsions. Glyceryl Stearate (GS) was mixed with Medium Chain Trygliceride (MCT) oil to formulate SLNs. SLNs presented a significantly enhanced ß-carotene retention and a slower ß-carotene degradation kinetics at increasing storage temperature, acidic conditions and light exposure. In fact, SLNs formulated with 5% GS in the lipid phase and stored at 4 °C and pH 7 retained almost 70% of the initially encapsulated ß-carotene after 55 days of storage, while it was completely degraded when it was encapsulated in liquid nanoemulsions. Moreover, it was observed that the solid lipid type affects the protective effect that SLNs may confer to the encapsulated lipophilic bioactives. Saturated long chain triglycerides, such as hydrogenated palm oil (HPO) presented slower and lower ß-carotene degradation kinetics in comparison to solid lipids composed of MCT, such as Coconut Oil (CNUT) or MCT + 5% of GS in the lipid phase. This work evidences that the incorporation of lipophilic bioactive compounds, such as ß-carotene, into SLNs slows down their degradation kinetics which might be attributed to a reduced diffusion of the oxidative species due to the lipid crystalline structure.

Phenolic profiling and in vitro bioactivity of Moringa oleifera leaves as affected by different extraction solvents.

In this work the (poly)-phenolic profile of Moringa oleifera leaves was comprehensively investigated through untargeted metabolomics, following a homogenizer-assisted extraction (HAE) using three solvent systems, i.e. methanol (HAE-1), methanol-water 50:50 v/v (HAE-2) and ethyl acetate (HAE-3). This approach allowed to putatively annotate 291 compounds, recording mainly flavonoids and phenolic acids. Thereafter, antioxidant capacity, antimicrobial activity and enzyme inhibition were assayed in the different extracts. HAE-1 extract showed the highest total phenolic content (31.84 mg/g), followed by HAE-2 (26.95 mg/g) and HAE-3 (14.71 mg/g). In addition, HAE-1 and HAE-2 extracts exhibited an expressive activity against Bacillus cereus and Listeria innocua. The HAE-2 leaf extract was characterized by the highest DPPH and ABTS values (being 49.55 and 45.26 mgTE/g), while ferric reducing antioxidant power was found to be higher in HAE-1 (58.26 mgTE/g). Finally, the enzyme inhibitory effects of M. oleifera leaf extracts were investigated against five enzymes, namely acetylcholinesterase (AChE), butyrylcholinesterase (BChE), tyrosinase, α-amylase and α-glucosidase. All of the tested extracts exhibited inhibitory effects on AChE and BChE with a higher activity for HAE-3 and HAE-1, whilst HAE-1 showed the higher impact on tyrosinase, glucosidase and amylase activities. Taken together, these findings suggest that M. oleifera leaf extracts are a good source of bioactive polyphenols with a potential use in food and pharma industries.

The lipid type affects the in vitro digestibility and ß-carotene bioaccessibility of liquid or solid lipid nanoparticles.

Solid lipid nanoparticles (SLNs) are emulsion-based carriers of lipophilic bioactive compounds. However, their digestibility may be affected by the solid lipid phase composition. Hence, the aim of this work was to study the in vitro lipolysis kinetics as well as the relationship between the lipid digestion, micelle fraction composition and ß-carotene bioaccessibility of SLNs with different solid lipids, being blends of medium chain triglyceride (MCT) oil, glyceryl stearate (GS) or hydrogenated palm oil (HPO) as compared to liquid lipid nanoparticles (LLNs) with pure MCT. SLNs formulated with GS were fully digested, similarly to LLNs. However, HPO-containing SLNs presented slower lipolysis kinetics during the intestinal phase at increasing HPO concentration. Despite this, HPO-SLNs showed higher ß-carotene bioaccessibility, which was related to the higher amount of monounstaturated free fatty acids in the micelle fraction. Thus, this work provides valuable insight for designing delivery systems of bioactive compounds with optimal functionality.