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Exopolysaccharides (EPSs) are important bioactive carbohydrate compound such as Capsular polysaccharide (CPS). EPSs are often used as bioenrichment agents and bioabsorbers to remove environmental pollutants like di-n-butyl phthalate (DBP). Among the EPSs producing bacteria, lactic acid bacteria (LAB) have gained the most attention. As generally recognized as safe (GRAS) microorganisms, LAB can produce EPSs with many different structures without any health risks. However, the EPSs production of LAB is usually not enough for large-scale application in industrial production. Here, capA gene (encoding CPS biosynthesis protein) was overexpressed in Lactobacillus plantarum P1 to improve the production of EPSs and further enhance the capability of the adsorption of DBP. Compared with P1, the CPS production in capA overexpressed strain was increased by 11.3 mg/L, and the EPSs thickness was increased from 0.0786±0.0224μm in P1 to 0.1160±0.0480μm in P1-capA. These caused the adsorption ratio of DBP in P1-capA to be doubled. Overall, the findings in this study provided a safe solution for the adsorption and removing of DBP.

Background: Lipid nanocarriers have been widely tested as drug delivery systems to treat diseases due to their bioavailability, controlled release, and low toxicity. For the pulmonary route, the Food and Drug Administration favors the use of substances generally recognized as safe, as well as biodegradable and biocompatible to minimize the possibility of toxicity. Tuberculosis (TB) remains a public health threat worldwide, mainly due to the long treatment duration and adverse effects. Therefore, new drug delivery systems for treating TB are needed. Objective: Physicochemical characterization of different lipid-based nanocarriers was used to optimize carrier properties. Optimized systems were incubated with Mycobacterium tuberculosis to assess whether lipid-based systems act as the energy source for the bacteria, which could be counterproductive to therapy. Method: Several excipients and surfactants were evaluated to prepare different types of nanocarriers using high-pressure homogenization. Results: A mixture of trimyristin with castor oil was chosen as the lipid matrix after differential scanning calorimetry analysis. A mixture of egg lecithin and PEG-660 stearate was selected as an optimal surfactant system, as this mixture formed the most stable formulations. Three types of lipid nanocarriers, solid lipid nanoparticles, nanostructured lipid carriers (NLC), and nanoemulsions, were prepared, with the NLC systems showing the most suitable properties for further evaluation. It may provide the advantages of increasing the entrapment efficiency, drug release, and the ability to be lyophilized, producing powder for pulmonary administration as an alternative to entrap poor water-soluble molecules. Conclusion: Furthermore, the NLC system can be considered for use as a platform for the treatment of TB through the pulmonary route.

Bacillus subtilis CW14, isolated from fresh elk droppings in Beijing Zoo, is a Gram-positive, conferred Generally Recognized as Safe (GRAS) bacterium with the capacity of ochratoxin A (OTA) detoxification. The genome sequence of the CW14 strain showed a size of 4,287,522 bp with 44.06% GC content. It was predicted many putative enzymes involved in degrading mycotoxin by analyzing the signal peptides and the transmembrane regions. Nine extracellular enzymes were predicted relating to OTA detoxification, including four D-Ala-D-Ala carboxypeptidases, two hydrolases, two amidases, and one lactamase. Indeed, two of the carboxypeptidase genes dacA and dacB, expressed in Escherichia coli, were verified contributing to OTA detoxification. DacA and OTA were mixed incubated for 24 h, and the degradation rate reached 71.3%. After purification, the concentration of recombinant DacA protein was 0.5 mg/mL. Bacillus subtilis CW14 and its carboxypeptidases may be used as OTA detoxification agents in food and feed industry production.

Purpose To date, a total of 13 probiotic Bacillus species are considered as a Generally Recognized as Safe organism (GRAS) approved by the US Federal Food, Drug, and Cosmetic Act (FDCA), which are used for food and feed additives. However, Bacillus velezensis is not considered as a probiotic candidate in swine farming due to a lack of genetic basis of probiotic action-related traits. Therefore, the present study was undertaken to exploit the genetic basis underlying the probiotic traits of B. velezensis VTX9. Methods The genome sequencing of B. velezensis VTX9 was performed on a PacBio Sequel platform. The probiotic properties including biosafety, antioxidative capacity, and riboflavin and exopolysaccharide production were evaluated by using genotypic and phenotypic analysis. The secondary metabolite potentials were also predicted. Results Strain VTX9 isolated from swine feces proved some probiotic properties including resistance to 3 mM H2O2, 0.6 mM bile salt, low pH, and antipathogenic activity. The complete genome of B. velezensis VTX9 consists of a 3,985,800 bp chromosome that housed 3736 protein-coding genes and 5 plasmids with the size ranging from 7261 to 20,007 bp. Genome analysis revealed no functional genes encoding enterotoxins and transferable antibiotic resistance, which confirmed the safety of VTX9. A total of 82 genes involved in gastrointestinal stress tolerance were predicted, which has not been reported previously. The maximum production of riboflavin reached 769 ± 7.5 ng/ml in LB medium after 72 h, which was in agreement with the complete de novo riboflavin biosynthetic pathway exploited for the first time in the B. velezensis genome. Antagonistic activity against pathogenic bacteria was attributed to 10 secondary metabolites clusters. The presence of a large gene cluster involved in biosynthesis of exopolysaccharides underscored further the adhesion and biofilm-forming capabilities of VTX9 in swine intestines. Conclusion Our results revealed for the first time that B. velezensis VTX9 has the potential to be a probiotic candidate. The information provided here on the genome of B. velezensis VTX9 opens new opportunities for using B. velezensis as a feed additive for swine farming in the future.

The biologically active phytochemicals are sourced from edible and medicinally important plants and are important molecules being used for the formulation of thousands of drugs. These phytochemicals have great benefits against many ailments particularly the inflammatory diseases or oxidative stress-mediated chronic diseases. Eugenol (EUG) is a versatile naturally occurring molecule as phenolic monoterpenoid and frequently found in essential oils in a wide range of plant species. EUG bears huge industrial applications particularly in pharmaceutics, dentistry, flavoring of foods, agriculture, and cosmeceutics. It is being focused recently due to its great potential in preventing several chronic conditions. The World Health Organization (WHO) has declared EUG as a nonmutant and generally recognized as safe (GRAS) molecule. The available literature about pharmacological activities of EUG shows remarkable anti-inflammatory, antioxidant, analgesic, and antimicrobial properties and has a significant effect on human health. The current manuscript summarizes the pharmacological characteristics of EUG and its potential health benefits.

Polyols are poorly digestible carbohydrates present in small amounts in some fruits and vegetables. Xylitol and erythritol are used as table sweeteners. These compounds are widely used in the food industry due to their low-calorie content. Erythritol is the only noncaloric polyol. Xylitol is the sweetest of the polyols, being the only one with sweetness equivalent to sucrose, but with one third of its calories. Clinical studies have shown reductions in the number of plaques, in counts of Streptococcus mutans, and in the number of dental cavities in individuals receiving erythritol and xylitol. Xylitol is also capable of reducing the growth and adherence to the oropharynx of bacteria that cause acute otitis media, such as Streptococcus pneumoniae, and several studies have shown that it reduces the risk of this bacterial infection in children. In addition to these effects, polyols can also have beneficial effects on metabolism. Both erythritol and xylitol have been approved by the European Union for use as sweeteners for several years, and replacing sugar with polyols decreases caloric intake, which can reduce body weight and blood glucose in individuals with obesity and type 2 diabetes mellitus. The safety of polyols is recognized by the Food and Drug Administration (FDA), who classifies them as compounds generally recognized as safe (GRAS). Thus, based on available scientific data, the daily consumption of both substances is associated with several benefits and does not represent any risk to human health.

It is widely known that an adequate intake of proteins with essential amino acids stimulates the production of essential nonprotein substances for the body, such as serotonin, thyroid hormones, and glutathione. Biotechnology applications can focus on food and its benefits for human health, either by improving the nutritional value of existing foods or developing new alternatives that can help resolve the effects of poor nutrition. The yeast Pichia pastoris has been designated as a GRAS (generally recognized as safe) microorganism and is currently used to produce a large number of recombinant proteins. In this study, a 100% synthetic protein was expressed intracellularly as a protein with high nutritional quality that contains a 75% ratio of essential amino acids using the expression system of the yeast P. pastoris. This approach might offer future possibilities for using the modified yeast as a single-cell protein source in the livestock industry and the food supplement market to reduce total protein intake.

Today’s well-informed consumer is demanding health promoting foods which not only have natural ingredients and are additive free, but also offer functional properties without compromising on the taste. Excessive consumption of sugar is a leading cause of non- communicable diseases, globally. Not more than 10% of the calories should be derived from sugar for optimal health. This has led to the increase in demand of food products containing sugar free alternatives. Therefore, naturally occurring non-nutritive sweeteners that have positive effects on the body weight as well as metabolism may facilitate in limiting the sugar intake and accomplishing the present recommendations. They are also an excellent alternative both for the consumers as well as the food industry in place of artificial sweeteners. These natural sweeteners are extracted from indigenous plants, provide insignificant calories, taste like sucrose, do not exhibit metallic after taste, and moreover, offer significant health benefits. The steviol glycosides and Luo Han Guo fruit (Monk fruit) are the natural extracts of plants, which are commercialized. In addition, numerous plant proteins such as thaumatin, brazzein, miraculin and several carbohydrates as rare sugars such as D- tagatose, D-allulose, D-sorbose and D-allose are being explored as potential substitutes to intense sweeteners which are reviewed in this paper. These are being extensively studied by the researchers for their physicochemical and functional properties and their possible use in low- calorie food formulations. They have a history of safe consumption by the natives of the country of their origin and have a generally recognized as safe (GRAS) or a novel food status.

Background The biodeterioration of food commodities by microbial pathogens remains a major public health concern. Worldwide, investigations have been carried out to develop harmless natural food preservatives based on essential oils (EOs) to protect food commodities, and advances have been made to meet users’ acceptance as a substitute for synthetic preservatives. Scope and approach In recent years, antimicrobial and antioxidant potential of the EOs from Thymus species have been documented with increasing demands from legislation changes, adaptation to the consumer trends, search for alternatives to solve rising food microbial pathogen resistance, and substitution of synthetic preservatives associated with adverse health effects. Hence, the present review paper emphasizes on the antimicrobial and antioxidant applications of EOs from different species of the genus Thymus and discloses the gaps where investigations are required. In addition, progress in the using Thymus EOs (TEOs) based nanoemulsions in food preservation, and their challenges in the application in food systems have also been discussed. Key findings and conclusions TEOs are rich in pharmacologically active constituents that confirm their industrial and health applications. In addition, TEOs and their nanoemulsions not only provide themselves to exploit in the food industry, but are also put under GRAS (Generally Recognized As Safe) category, and have shown potential inhibitory activity against a broad range of pathogens in food commodities. Thus, TEOs can be a source for the development of natural preservatives that meet the needs of the food industry to satisfy both its requirements and those of the consumers. However, despite the potential for EOs, more studies are required to assess their probable side effects and safety levels before considering their deployment for food purposes.

The treatments being investigated for the pandemic associated with a severe acute respiratory syndrome caused by the new coronavirus (SARS-CoV2) aim to inhibit viral replication or inhibit enzymatic elastase in the lung, responsible for the SARS-CoV cell entry mechanism and the pathogenesis of severe pneumonia caused by infection There is also a need to improve alveolar surfactant capacity and inhibition of the exacerbated inflammatory response Current knowledge allows us to infer that Citrus essential oils could be considered an individual or a complementary highly valuable alternative for the viral infection caused by SARS-CoV due to its antiviral activity, particularly against coronavirus, anti-elastase, anti-inflammatory, antibacterial and antibiofilm properties Furthermore, its proven capacity of biosurfactants, antioxidants, and pain inhibitors would help to treat acute respiratory syndrome This review aims to encourage the development of more detailed studies on Citrus essential oils as a new multi-objective strategy to treat SARS-CoV2 infection Abbreviations: SARS-CoV: Severe Acute Respiratory Syndrome Coronavirus;GRAS: Generally Recognized as Safe;TRP: Transient Receptor Proteins;HeLa-CEACAM1a: HeLa-epithelial carcinoembryonic antigen-related cell adhesion molecule 1a;HSV: Herpes Simplex Virus;FDA: Food and Drug Administration;ANMAT: National Administration of Medicines, Food and Medical Technology, Argentina;AFSSAPS: Agence Francaise de Securite Sanitaire des Produits de Sante, France;ACE2:Angiotensin-converting enzyme-2 [ABSTRACT FROM AUTHOR] Copyright of Journal of Essential Oil Research is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission However, users may print, download, or email articles for individual use This abstract may be abridged No warranty is given about the accuracy of the copy Users should refer to the original published version of the material for the full abstract (Copyright applies to all Abstracts )

Eyelid plays a vital role in protecting the eye from injury or infection. Inflammation related eyelid diseases, such as blepharitis, are the most common ocular disorders that affect human's vision and quality of life. Due to the physiological barriers and anatomical structures of the eye, the bioavailability of topical administrated therapeutics is typically less than 5%. Herein, we developed a bio-responsive hydrogel drug delivery system using a generally recognized as safe compound, triglycerol monostearate (TG-18), for in-situ eyelid injection with sustained therapeutics release. In vitro, drug release and disassembly time of Rosiglitazone loaded hydrogel (Rosi-hydrogel) were estimated in the presence or absence of MMP-9, respectively. Moreover, the disassembly of TG-18 hydrogel was evaluated with 9-month-old and 12-month-old mice in vivo. Owing to the bio-responsive nature of Rosi-hydrogel, the on-demand Rosiglitazone release is achieved in response to local enzymes. These findings are proved by further evaluation in the age-related meibomian gland dysfunction mice model, and the bio-responsive hydrogel is used as an in-situ injection to treat eyelid diseases. Taken together, the in-situ eyelid injection with sustained drug release opens a window for the therapy of inflammation related eyelid diseases., Highlights • This study is the first application of injectable bio-responsive hydrogel for therapy of inflammation related eyelid diseases. • The enzyme response characteristic is extremely suitable for enhancing drug bioavailability in ocular drug delivery. • In-situ release of rosiglitazone can effectively treat age-related meibomian gland dysfunction in the mice model.

Gamma-aminobutyric acid (GABA) is an important non-protein amino acid with wide-ranging applications. Currently, GABA can be produced by a variety of methods, including chemical synthesis, plant enrichment, enzymatic methods, and microbial production. Among these methods, microbial production has gained increasing attention to meet the strict requirements of an additive in the fields of food, pharmaceutical, and livestock. In addition, renewable and abundant resources, such as glucose and lignocellulosic biomass can also be used for GABA microbial production under mild and environmentally friendly processing conditions. In this review, the applications, metabolic pathways and physiological functions of GABA in different microorganisms were firstly discussed. A comprehensive overview of the current status of process engineering strategies for enhanced GABA production, including fermentation optimization and whole-cell conversion from different feedstocks by various host strains is also provided. We also presented the state-of-the-art achievements in strain development strategies for industrial lactic acid bacteria (LAB), Corynebacterium glutamicum and Escherichia coli to enhance the performance of GABA bioproduction. In order to use bio-based GABA in the fields of food and pharmaceutical, some Generally Recognized as Safe (GRAS) strains such as LAB and C. glutamicum will be the promising chassis hosts. Toward the end of this review, current challenges and valuable research directions/strategies on the improvements of process and strain engineering for economic microbial production of GABA are also suggested.

Parabens are chemicals containing alkyl-esters of p-hydroxybenzoic acid, which give them antimicrobial, antifungal, and preservative properties. Propylparaben (PP) is one paraben that has been widely used in personal care products, cosmetics, pharmaceuticals, and food. In this review, we address the ongoing controversy over the safety of parabens, and PP specifically. These chemicals have received significant public attention after studies published almost 20 years ago suggested plausible associations between PP exposures and breast cancer. Here, we use key characteristics, a systematic approach to evaluate the endocrine disrupting properties of PP based on features of “known” endocrine disruptors, and consider whether its classification as a “weak” estrogen should alleviate public health concerns over human exposures. We also review the available evidence from rodent and human studies to illustrate how the large data gaps that exist in hazard assessments raise concerns about current evaluations by regulatory agencies that PP use is safe. Finally, we address the circular logic that is used to suggest that because PP has been used for several decades, it must be safe. We conclude that inadequate evidence has been provided for the safe use of PP in food, cosmetics, and consumer products.

Polyglycerol monolaurates are generally recognized as safe food additives and are commonly used as food emulsifiers. In this study, the antimicrobial effect of four polyglycerol monolaurates on two Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) and two Gram-negative bacteria (Escherichia. coli and Pseudomonas aeruginosa) were investigated. The minimum inhibitory concentration (MIC) of diglycerol monolaurate (PG2ML), triglycerol monolaurate (PG3ML), hexaglycerol monolaurate (PG6ML), and decaglycerol monolaurate (PG10ML) against S. aureus was 0.16, 0.32, 0.63, and 1.25 mg/mL, respectively. The MIC of PG2ML, PG3ML, PG6ML, and PG10ML against B. subtilis was 0.32, 0.63, 1.25, and 3.75 mg/mL, respectively. No apparent antimicrobial effect of these four polyglycerol monolaurates on E. coli and P. aeruginosa was observed even up to 10.00 mg/mL. The underlying mechanism was investigated by assessing cell membrane permeability, the integrity of cell membrane, and morphology. We concluded that polyglycerol monolaurates might eliminate Gram-positive bacteria by disrupting the cell membrane, thereby increasing cell membrane permeability, releasing the cellular contents, and altering the cell morphology.

Here, we report a novel, “dry”, nano-aerosol-based, antimicrobial technology using engineered water nanostructures (EWNSs) for leafy vegetable disinfection. These EWNS based nano-sanitizers are synthesized using a combined process of electrospray and ionization of aqueous solutions of generally recognized as safe active ingredients (AIs). A novel application of the EWNS to vegetable surfaces was carried out in this study. The EWNS-based nano-sanitizers were synthesized using an AI cocktail (10% hydrogen peroxide, 1% citric acid, 0.1% lysozyme and 0.0025% nisin), and their efficacy against foodborne pathogen surrogates and plant pathogens was assessed using a leafy vegetable model, namely spinach (Spinacia oleracea). The EWNSs were directed onto a coupon cut from spinach leaves, which were inoculated with microorganisms relevant to food safety, such as E. coli. It was shown that a 2 min exposure to EWNS-based nano-sanitizers significantly (P 0.05) in color or surface pH of spinach was found after 15 minutes of exposure. In summary, EWNS based nano-sanitizers are highly effective against foodborne pathogen surrogates and plant pathogens using only nanograms of nature-derived active ingredients and without an impact on produce quality.

A compound K-producing fungus was isolated from meju (fermented soybean brick) and identified as the generally recognized as safe (GRAS) strain Aspergillus tubingensis. The extracellular enzymes obtained after the cultivation of 6 days in the medium with 20 g/L citrus pectin as an inducer showed the highest compound K-producing activity among the inducers tested. Under the optimized conditions of 0.05 mM MgSO4, 55 °C, pH 4.0, 13.4 mM protopanaxadiol (PPD)-type ginsenosides, and 11 mg/mL enzymes, the extracellular enzymes from A. tubingensis completely converted PPD-type ginsenosides in the ginseng extract to 13.4 mM (8.35 mg/mL) compound K after 20 h, with the highest concentration and productivity among the results reported so far. As far as we know, this is the first GRAS enzyme to completely convert all PPD-type ginsenosides to compound K.

New strategies to efficiently treat bacterial infections are crucial to circumvent the increase of resistant strains and to mitigate side effects during treatment. Skin and soft tissue infections represent one of the areas suffering the most from these resistant strains. We developed a new drug delivery system composed of the green algae, Chlamydomonas reinhardtii, which is generally recognized as safe, to target specifically skin diseases. A two‐step functionalization strategy was used to chemically modify the algae with the antibiotic vancomycin. Chlamydomonas reinhardtii was found to mask vancomycin and the insertion of a photocleavable linker was used for the release of the antibiotic. This living drug carrier was evaluated in presence of Bacillus subtilis and, only upon UVA1‐mediated release, growth inhibition of bacteria was observed. These results represent one of the first examples of a living organism used as a drug delivery system for the release of an antibiotic by UVA1‐irradiation., Algae control the release: A new approach for antibiotic caging and controlled release was developed using Chlamydomonas reinhardtii. The algae were covalently modified with vancomycin, and using a photocleavable linker, light‐mediated release of an antibacterial agent from the cell surface and inhibited the growth of Bacillus subtilis (see figure).

In 2015, the US Food and Drug Administration passed a ban on the “generally recognized as safe” status of partially hydrogenated oils (PHOs), and in June 2018, PHOs were prohibited from being used. Our objective was to develop a predictive model to quantify trans-fat concentrations in bakery and snacks products using a portable mid-infrared (MIR) spectrometer. The approach was tested using 24 calibration standards (consisting of trielaidin in triolein and tripalmitin) and 87 bakery and snack products ranging from ND to 65% trans-fat. The fat was extracted by grinding products into powders and extracting the fat using petroleum ether. Gas Chromatography (AOCS Cd 14c-94) was used to determine the fatty acid profile and trans-fat content. Spectra were acquired by directly placing the fat (200 μL) onto the heated (65 ± 1°C) 5–reflection ZnSe crystal of a portable MIR spectrometer. Partial least squares regression (PLSR) models were developed using the calibration standards and extracted fats spectra targeting the signal of the C–H out-of-plane deformation band at 966 cm−1. Best model performances were obtained using the spectra of the extracted fat from bakery and snack products with the standard error of prediction of 0.5 g of trans-fats per 100 g of fat. We found that 25% of products labeled as zero trans-fat/serving did not comply with the maximum tolerance levels based on GC-FAME analysis. Portable FTIR devices operating in attenuated total reflection (ATR) mode can provide the food industry and government food inspectors with rapid, accurate, and high throughput measurements for routine screening to facilitate regulatory compliance.

Some additives used in polymeric products can cause health problems; thus, lowering migration of such substances from the polymeric matrix is of concern. Given that migration is sometimes inevitable, the use of safe additives is an alternative. This work demonstrates the viability of employing generally recognized as safe (GRAS) substances (citric acid, vitamin C, and vitamin E) on PVC films plasticized with epoxidized waste cooking oil. The prepared films presented good mechanical properties, and in a few cases even exhibited improvements in properties such as increase in tensile strength and decrease in water vapor permeability. The films promoted a significant reduction in browning of refrigerated apple samples. The vitamin C-laden film was the most effective, being able to diminish apple browning even at room temperature. Among the tested additives, citric acid was the most effective in terms of protective action against oxidation, preventing degradation of procyanidin B2, caffeic acid, and epicatechin.

We determined the effects of a commercially available, GRAS (Generally Recognized As Safe) by independent conclusion, CBD-containing hemp oil extract on stress resilience, perceived recovery, mood, affect, body composition, and clinical safety markers in healthy human subjects.

In this study, the effect of three essential oils (EOs) – clove oil (CO), thyme oil (TO), and garlic oil (GO), which are generally recognized as safe – on the planktonic growth, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), motility, biofilm formation, and quorum sensing (QS) of Vibrio parahaemolyticus was investigated. All three EOs showed bacteriostatic activity, with MICs in the range 0.02%–0.09% (v/v). CO and TO completely controlled planktonic growth at 0.28% and 0.08% (v/v), which is four times their MIC (4 × MIC), after 10 min, whereas GO completely controlled growth at 0.36% (v/v) (4 × MIC) after treatment for 20 min. V. parahaemolyticus motility was significantly reduced by all three EOs at 4 × MIC (0.28% for CO, 0.08% for TO, and 0.36% for GO), whereas QS was controlled and biofilm formation reduced by all three EOs at 8 × MIC (0.56% for CO, 0.16% for TO, and 0.72% for GO) after 30 min of treatment. These results suggest that CO, TO, and GO have a significant inhibitory effect on V. parahaemolyticus cells in biofilm sand thus represent a promising strategy for improving food safety. These results provide the evidence required to encourage further research into the practical use of the proposed EOs in food preparation processes.

Background and objective Lactic acid bacteria are generally recognized as safe that could be beneficial for several uses in food industry to get their health benefits. The present study was focused on the isolation and identification of some new lactic acid bacteria that might be naturally occurred in the honeybees stomach and tried to explore their benefits. Materials and methods Twenty five isolates of lactic acid bacteria were isolated from the stomach of three different types of Egyptian bees (Apis mellifera lamarckii ), Carniolan bees (A.m. carnica) and hybrid Carniolan bees. Identification of isolates was carried out based on phenotypical tests and carbohydrate assimilation using API50 CHL and 16S rDNA sequencing. Results In the present study, the results emphasized Lactobacillus plantarum to be the predominant species (62.5%), other strains were identified as Pediococcus pentosaceus (12.5%), Lactobacillus pentosus (12.5%) and Lactobacillus sakei (12.5%). Eight of 25 isolates showed a potential antibacterial activity especially against Salmonella senftenberg strain. The novel isolates (HBMSS1, HBMSS3, HBMSS4, HBMSS5, HBMSS6 and HBMSS8) showed a significant antimicrobial activity against C. botulinum, E. coli, S. Senftenberg and S. epidermidis as food borne pathogens and P. larvae and M. plutonius as honeybee pathogens. Conclusion These promising findings might be beneficial for discovering novel preservatives in food industry and substitution of antibiotic drugs used in the treatment of honeybees' infection.

Background Phytic acid (Phytate) present in plant-based foods is an anti-nutrient because of its tendency to chelate various nutrients thus reducing their bioavailability in animal and human physiological system. Use of phytases which bring sequential cleavage of phosphate groups from phytic acid-mineral complexes, liberating minerals in solubilized form is a viable alternative. There are many reports on phytases of different microbial origin but due to safety aspects their use in food applications is debatable. Phytases from Lactic Acid Bacteria (LAB) which are generally recognized as safe have more scope for their application in food processing. Scope and approach LAB are among the key safe bacteria which are known to give probiotic and other health benefits. Therefore, there is a growing interest in the exploration of phytase producing LAB and their use in food processing. Number of reports are there on phytases from LAB and their applications. To further increase the scope in this area comprehensive information is required. Key findings and conclusions This review summarizes the various reports on LAB phytases with respect to their production, characterization, application in processing of various types of food and heterologous expression. Limitations and future scope of LAB phytases have also been discussed which will open new prospects of research for their application.

Nuts contain a vast array of phenolic compounds having important biological properties. They include substances allocated into the five major groups named phenolic acids, flavonoids, tannins, phenolic lignans, and stilbene derivatives. The complexity in composition does not allow for setting a universal extraction procedure suitable for extraction of all nut phenolics. The use of non-conventional extraction techniques, such as those based on microwave, ultrasound, and compressed fluids, combined with generally recognized as safe solvents is gaining major interest. With regard to the latter, ethanol, water, and ethanol-water mixtures have proven to be effective as extracting solvents and allow for clean, safe, and low-cost extraction operations. In recent years, there has been an increasing interest in biological properties of natural phenolic compounds, especially on their role in the prevention of several diseases in which oxidative stress reactions are involved. This review provides an updated and comprehensive overview on nut phenolic extraction and their chemical profiles and bioactive properties. Fil: Bodoira, Romina Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina Fil: Maestri, Damian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina

Food contamination by fungi and mycotoxins presents a problem for food safety even today. Since lactic acid (LA) has Generally Recognized As Safe (GRAS) status, the aim of this research was to determine its potential in protection of food against mycological and mycotoxicological contamination. In this study, LA showed an inhibitory effect on the growth of food-borne fungi (Penicillium aurantiogriseum K51, Aspergillus parasiticus KB31, Aspergillus versicolor S72, and Aspergillus niger K95) and on biosynthesis of sterigmatocystin (STE). For the antifungal effect of LA on the growth of food-borne fungi, the disc diffusion and microdilution methods were performed. The effect of LA on the STE biosynthesis by A. versicolor was determined using an LC-MS/MS technique. The largest inhibition zone was observed for A. versicolor (inhibition zone of 24 ± 0.35 mm), while there were no inhibition zones for A. niger and A. parasiticus at all tested LA concentrations. The minimal inhibitory concentration (MIC) of LA on fungi ranged from 25.0 mg/mL to 50.0 mg/mL, while the minimum fungicidal concentrations (MFCs) ranged from 50.0 mg/mL to 100.0 mg/mL. Complete inhibition of STE biosynthesis by A. versicolor was observed at an LA concentration of 50.0 mg/mL. The obtained results showed that LA could be efficient for protection of food against mycological and STE contamination.

l-threonine, l-tryptophan and l-valine play a fundamental role in animal and human nutrition as essential amino acids required for normal growth. In addition, each amino acid is codified as a generally recognized as safe (GRAS) amino acid for the use in animal feed additives and presents no exposure risk from animal to humans consuming tissues or products from the target animal. Taking into account the important role of mutagenicity and genotoxicity in the risk of the three amino acid additives (l-threonine, l-tryptophan, and l-valine) fermentation products and other unknown impurities and derivatives from Corynebacterium glutamicum (C. glutamicum), the safety evaluation of these amino acid additives is not performed. Therefore, the purpose of this study is to evaluate toxicological effects, including Ames test, an in vitro mammalian chromosomal aberration test and an acute oral animal toxicity of the three amino acid additives in accordance with the Organisation for Economic Co-operation and Development (OECD) guidelines and the principles of Good Laboratory Practice (GLP). As a result, these amino acid additives were classified as non-mutagenic and non-clastogenic, and did not induce any toxicity in acute oral toxicity test. Collectively, these results suggest that the three amino acid additives are safe with no adverse effects, and able to be applied as an ingredient or other biological uses.

Rhizopus oryzae is a fungus used to ferment tempeh in Indonesia and is generally recognized as safe (GRAS) for human consumption by the USA FDA. We previously assessed the effect of a tempeh extract on cortisol levels in zebrafish but did not include behavioral studies. Here, we measured the GABA content in three strains of Rhizopus oryzae, two isolated by us (MHU 001 and MHU 002) and one purchased. We then investigated the effect of tempeh on cortisol and the gut microbiota in a zebrafish experimental model. GABA concentration was the highest in MHU 002 (9.712 ± 0.404 g kg−1) followed by our MHU 001 strain and the purchased one. The fish were divided into one control group fed a normal diet and three experimental groups fed soybean tempeh fermented with one of the three strains of Rhizopus oryzae. After two weeks, individual fish were subjected to unpredicted chronic stress using the novel tank diving test and the tank light–dark test. Next-generation sequencing was used to analyze gut microbial communities and RT-PCR to analyze the expression of BDNF (brain-derived neurotrophic factor) gene and of other genes involved in serotonin signaling/metabolism in gut and brain. Tempeh-fed zebrafish exhibited increased exploratory behavior (less stress) in both tank tests. They also had significantly reduced gut Proteobacteria (include E. coli) (51.90% vs. 84.97%) and significantly increased gut Actinobacteria (include Bifidobacterium spp.) (1.80% vs. 0.79%). The content of Bifidobacteriumadolescentis, a “psychobiotic”, increased ten-fold from 0.04% to 0.45%. Tempeh also increases BDNF levels in zebrafish brain. Rhizopus oryzae MHU 001 greatly improved the anti-stress effect of tempeh and microbiota composition in zebrafish gut.

Joint review with authors from Universiti Putra Malaysia, Malaysia Food Research Unit, Department of Agriculture, Sri Lanka De Montfort University, United Kingdom University of Tikrit, Tikrit, Iraq Mango (Mangifera indica L.) is one of the most important tropical fruits due to its pleasant taste and high nutrient content including vitamins, minerals, and phytochemicals. Post-harvest fungal diseases, mainly anthracnose, stem-end rot, and aspergillus rot are among the major problems in mango provision throughout all the supply chain stages. Fungal diseases create large economic losses and present a health risk for consumers due to the production of mycotoxins (aflatoxin and patulin). In the last two decades, chemical methods for fungal control have been prevalent because of their low cost and simple practicing features. However, recent consumer trends and regulations have limited the use of chemicals in fruits at the post-harvest stage. Bio-preservation is a promising alternative technique applied to control the post-harvest fungi with high safety for the consumer and environment. Antifungal polypeptides are made by different living beings including bacteria, fungi, protozoa, plants, invertebrates, and vertebrates. Polypeptides including small peptides produced by lactic acid bacteria (LAB) have a higher potential to employ as food bio-preservatives due to their promising antifungal ability with generally recognized as safe (GRAS) status. Lacto-fermented polypeptides inhibit a broad range of fungal growth, and thus enhance the quality and extend the shelf life through reducing biotic stress, respiration, and ethylene biosynthesis of post-harvested fruits. Recently, polypeptides, mainly bioactive peptides, and bacteriocins received great attention for their potential bio-preservation activity against pathogenic fungi in fruits. Moreover, the development of edible coating enhanced the stability of the bioactive polypeptides and expanded their applications for fruit preservation. This article reviews the fungi associated with main mango diseases, major causal organisms, antifungal activity of LABs’ polypeptides on mango, and their antifungal performances to control the postharvest mango spoilage. Besides, this review highlights the potential applications of Lacto-fermented polypeptides-enriched edible coating to control the pathogenic fungi of mango.

Antibiotics played an important role in controlling the development of enteric infection. However, the emergence of antibiotic resistance and gut dysbiosis led to a growing interest in the use of natural antimicrobial agents as alternatives for therapy and disinfection. Chitosan is a nontoxic natural antimicrobial polymer and is approved by GRAS (Generally Recognized as Safe by the United States Food and Drug Administration). Chitosan and chitosan derivatives can kill microbes by neutralizing negative charges on the microbial surface. Besides, chemical modifications give chitosan derivatives better water solubility and antimicrobial property. This review gives an overview of the preparation of chitosan, its derivatives, and the conjugates with other polymers and nanoparticles with better antimicrobial properties, explains the direct and indirect mechanisms of action of chitosan, and summarizes current treatment for enteric infections as well as the role of chitosan and chitosan derivatives in the antimicrobial agents in enteric infections. Finally, we suggested future directions for further research to improve the treatment of enteric infections and to develop more useful chitosan derivatives and conjugates.

Viral infections may cause serious human diseases. For instance, the recent appearance of the novel virus, SARS-CoV-2, causing COVID-19, has spread globally and is a serious public health concern. The consumption of healthy, proper, functional, and nutrient-rich foods has an important role in enhancing an individual's immune system and preventing viral infections. Several polysaccharides from natural sources such as algae, bacteria, and fungi have been considered as generally recognized as safe (GRAS) by the US Food and Drug Administration. They are safe, low-toxicity, biodegradable, and have biological activities. In this review, the bioactive polysaccharides derived from various microorganisms, including bacteria, fungi, and algae were evaluated. Antiviral mechanisms of these polysaccharides were discussed. Finally, the potential use of microbial and algal polysaccharides as an antiviral and immune boosting strategy was addressed. The microbial polysaccharides exhibited several bioactivities, including antioxidant, anti-inflammatory, antimicrobial, antitumor, and immunomodulatory activities. Some microbes are able to produce sulfated polysaccharides, which are well-known to exert a board spectrum of biological activities, especially antiviral properties. Microbial polysaccharide can inhibit various viruses using different mechanisms. Furthermore, these microbial polysaccharides are also able to modulate immune responses to prevent and/or inhibit virus infections. There are many molecular factors influencing their bioactivities, e.g., functional groups, conformations, compositions, and molecular weight. At this stage of development, microbial polysaccharides will be used as adjuvants, nutrient supplements, and for drug delivery to prevent several virus infections, especially SARS-CoV-2 infection.

Since antiquity, the ubiquitous lactic acid bacteria (LAB) Enterococci, which are just as predominant in both human and animal intestinal commensal flora, have been used (and still are) as probiotics in food and feed production. Their qualities encounter several hurdles, particularly in terms of the array of virulence determinants, reflecting a notorious reputation that nearly prevents their use as probiotics. Additionally, representatives of the Enterococcus spp. genus showed intrinsic resistance to several antimicrobial agents, and flexibility to acquire resistance determinants encoded on a broad array of conjugative plasmids, transposons, and bacteriophages. The presence of such pathogenic aspects among some species represents a critical barrier compromising their use as probiotics in food. Thus, the genus neither has Generally Recognized as Safe (GRAS) status nor has it been included in the Qualified Presumption of Safety (QPS) list implying drastic legislation towards these microorganisms. To date, the knowledge of the virulence factors and the genetic structure of foodborne enterococcal strains is rather limited. Although enterococcal infections originating from food have never been reported, the consumption of food carrying virulence enterococci seems to be a risky path of transfer, and hence, it renders them poor choices as probiotics. Auspiciously, enterococcal virulence factors seem to be strain specific suggesting that clinical isolates carry much more determinants that food isolates. The latter remain widely susceptible to clinically relevant antibiotics and subsequently, have a lower potential for pathogenicity. In terms of the ideal enterococcal candidate, selected strains deemed for use in foods should not possess any virulence genes and should be susceptible to clinically relevant antibiotics. Overall, implementation of an appropriate risk/benefit analysis, in addition to the case-by-case assessment, the establishment of a strain’s innocuity, and consideration for relevant guidelines, legislation, and regulatory aspects surrounding functional food development seem to be the crucial elements for industries, health-staff and consumers to accept enterococci, like other LAB, as important candidates for useful and beneficial applications in food industry and food biotechnology. The present review aims at shedding light on the world of hurdles and limitations that hampers the Enterococcus spp. genus and its representatives from being used or proposed for use as probiotics. The future of enterococci use as probiotics and legislation in this field are also discussed.

Serpa is an ovine raw milk cheese widely appreciated by the consumers. However, raw milk products may be seen with reservations in terms of safety or technological defects. To overcome that, the addition of an autochthonous starter culture may ensure the cheesemaking process optimization and microbiota dominance. In this work, the technological and protective performance of eleven lactic acid bacteria strains, isolated from Serpa Protected Designation of Origin cheese and reported as generally recognized as safe, were screened. The integration of technological and protective properties studied in the PCA plot, coupled with the proteolytic and lipolytic analysis suggested that Lb. plantarum PL1 and PL4 strains may be the best candidates. These strains showed both proteolytic and lipolytic activities, a good acidification potential, low d -lactic acid production and were well adapted to the salt and temperatures used. PL1 strain also exhibited a higher antimicrobial effect against the pathogenic bacteria studied. Although Lb. paracasei strain showed lower acidification capacity, due to their technological and protective properties, it could be combined with other more acidifying strains. As future work, it is important to establish cheese model systems to complement this screening and implement an autochthonous starter culture.

Microbial transglutaminase (mTG) is a heavily used food additive and its industrial transamidated complexes usage is rising rapidly. It was classified as a processing aid and was granted the GRAS (generally recognized as safe) definition, thus escaping full and thorough toxic and safety evaluations. Despite the manufacturers claims, mTG or its cross-linked compounds are immunogenic, pathogenic, proinflammatory, allergenic and toxic, and pose a risk to public health. The enzyme is a member of the transglutaminase family and imitates the posttranslational modification of gluten, by the tissue transglutaminase, which is the autoantigen of celiac disease. The deamidated and transamidated gliadin peptides lose their tolerance and induce the gluten enteropathy. Microbial transglutaminase and its complexes increase intestinal permeability, suppresses enteric protective pathways, enhances microbial growth and gliadin peptide’s epithelial uptake and can transcytose intra-enterocytically to face the sub-epithelial immune cells. The present review updates on the potentially detrimental side effects of mTG, aiming to interest the scientific community, induce food regulatory authorities’ debates on its safety, and protect the public from the mTG unwanted effects.

Lactic acid bacteria (LAB) are important members in synthetic microbial consortia due to their ‘generally recognized as safe’ status and diverse metabolic activities. Defined communities with LAB show great potential in elucidating metabolic interactions that drive their assembly and demonstrating power to address sustainability challenges in food, environment, and health.

Probiotics have been reported to play a major role in maintaining the balance of microbiota in host. Consumption of food with probiotics has increased with consumer concerns regarding healthy diets and wellness. Correspondingly, safety evaluation of probiotics for human consumption has become increasingly important in food industry. Herein, we aimed to test the safety of Bifidobacterium lactis BL-99 and Lacticaseibacillus paracasei K56 and ET-22 strains in vitro and in vivo. In results, these strains were found to be negative for mucin degradation and platelet aggregation test. Additionally, the three strains were susceptible to eight antibiotics. In accordance with bacterial reversion mutation (Ames) assay, the tested strains had no genetic mutagenicity. Finally, it was confirmed that there were no dose-dependent mortality and toxicity throughout multidose oral toxicity tests in rats. Our findings demonstrated that B. lactis BL-99 and L. paracasei K56 and ET-22 can achieve the generally recognized as safe (GRAS) status as probiotics in the future.

One of the most challenging problems with food-borne bioactive compounds is that there are commonly no cost-effective, generally recognized as safe (GRAS) methods for obtaining gram quantities of their purified forms. Here we aimed at developing a method to isolate deflamin, an oligomeric protein from lupin seeds with anti-inflammatory and anticancer activity through matrix metalloprotease (MMP)-9 inhibition. Our goal was to develop a GRAS method that could be easily up-scalable whilst maintaining deflamin’s activity. A sequential precipitation methodology was developed, using an aqueous extraction, followed by heat denaturation, acid precipitation and solubilization in ethanol. A final precipitation with 90% ethanol yielded a purified protein which was sequenced through mass spectrometry and tested for its MMP inhibitory activity using the Dyequenched (DQ) gelatin assay and the standard wound healing assay in HT29 cells. The developed method yielded a purified oligomer, which represented 0.1% (w/w) of total dry seed weight and was positively confirmed to be deflamin. It further showed to effectively reduce MMP-9 gelatinolytic activity as well as colon cancer cell migration, hence corroborating the effectiveness of our method. Overall, this is the first reported method for isolating an MMP-9 inhibitor from legume seeds, which is up-scalable to an industrial level, in a cost-effective manner info:eu-repo/semantics/publishedVersion

International audience; Cereals are subject to contamination by pathogenic fungi, which damage grains and threaten public health with their mycotoxins. Fusarium graminearum and its mycotoxins, trichothecenes B (TCTBs), are especially targeted in this study. Recently, the increased public and political awareness concerning environmental issues tends to limit the use of traditional fungicides against these pathogens in favor of eco-friendlier alternatives. This study focuses on the development of biofungicides based on the encapsulation of a curcumin derivative, tetrahydrocurcumin (THC), in polysaccharide matrices. Starch octenylsuccinate (OSA-starch) and chitosan have been chosen since they are generally recognized as safe. THC has been successfully trapped into particles obtained through a spray-drying or freeze-drying processes. The particles present different properties, as revealed by visual observations and scanning electron microscopy. They are also different in terms of the amount and the release of encapsulated THC. Although freeze-dried OSA-starch has better trapped THC, it seems less able to protect the phenolic compound than spray-dried particles. Chitosan particles, both spray-dried and lyophilized, have shown promising antifungal properties. The IC50 of THC-loaded spray-dried chitosan particles is as low as 0.6 ± 0.3 g/L. These particles have also significantly decreased the accumulation of TCTBs by 39%.