Generally Recognized As Safe Research Articles

Investigating the Antibacterial Effect of a Novel Gallic Acid-Based Green Sanitizer Formulation.

The purpose of the present study was to investigate the mechanism of action of our newly developed green sanitizer formulation comprising a natural phenolic compound, gallic acid (GA), strengthened by the Generally Recognized as Safe (GRAS) materials hydrogen peroxide (H2O2) and DL-lactic acid (LA). Combining 8 mM GA with 1 mM H2O2 resulted in an abundant generation of reactive oxygen species (ROS) and a bactericidal effect towards Gram-negative (Escherichia coli, Pseudomonas syringae, and Pectobacterium brasiliense) and Gram-positive (Bacillus subtilis) bacteria (4 to 8 log CFU mL-1 reduction). However, the exposure to this dual formulation (DF) caused only a modest 0.7 log CFU mL-1 reduction in the Gram-positive L. innocua population. Amending the DF with 20 mM LA to yield a triple formulation (TF) resulted in the efficient synergistic control of L. innocua proliferation without increasing ROS production. Despite the inability to grow on plates (>7 log CFU mL-1 population reduction), the TF-exposed L. innocua maintained high intracellular ATP pools and stable membrane integrity. The response of L. innocua to TF could be qualified as a "viable but nonculturable" (VBNC) phenomenon, while with the other species tested this formulation caused cell death. This research system may offer a platform for exploring the VBNC phenomenon, a critical food safety topic.

Extraction of Soybean Oil with Pressurized Ethanol: Prospects for a New Processing Approach with an Analysis of the Physical Properties of Crude Oil and Implementation Costs through Scale-Up in an Intermittent Process

New environmentally friendly methods for extracting vegetable oils are in development, with a focus on pressurized liquid extraction (PLE) in an intermittent process. Ethanol, a renewable and generally recognized as safe (GRAS) solvent, is gaining prominence in this process. It is crucial for these methods to maintain the physicochemical characteristics of the extracted oils and be economically viable on a large scale. Using SuperPro Design software v 8.5, a simulation of PLE scaled up to industrial levels was conducted. Measurements of oils extracted with pressurized hexane and ethanol showed minimal density variations, with slightly higher viscosity for ethanol-extracted oil. Accelerated oxidative degradation revealed a longer induction period for hexane-extracted oil, indicating that ethanol-extracted oil degrades more easily. The antioxidant activity of the oil extracted with hexane was lower than that with ethanol. In the differential scanning calorimetry analysis, the oils extracted with hexane and ethanol presented onset melting point temperatures of −43.2 and −36.1 °C, respectively. The economic assessment considered 16 scenarios, showing a return on investment ranging from 9.0 to 133.5% in the first year and payback times from 0.7 to 11.1 years. Scenario 8, involving two 5000 L extractors, ethanol recycling, and an annual production of 3,325,300 L of soybean oil at USD 1.25/L, demonstrated the best return on investment (133.5%) in less than one year. Overall, this study suggests that industrial-scale soybean oil extraction via PLE in an intermittent process can be more cost-effective than conventional methods, making implementation feasible.

Safety assessment of enterocin-producing Enterococcus strains isolated from sheep and goat colostrum

BackgroundThis study investigates the safety evaluation of enterocin-producing 11 E. mundtii and two E. faecium strains previously isolated from small livestock colostrums. Enterococcus species do not possess Generally Recognized as Safe (GRAS) status. Hence, it is critical to scrutinize enterococci’s antibiotic resistance, virulence characteristics, and biogenic amine production capabilities in order to assess their safety before using them as starter or adjunct cultures.ResultsEnterococcus strains showed susceptibility to medically significant antibiotics. Multiple-drug resistance (MDR) was found in only E. faecium HC121.4, and its multiple antibiotic resistance (MAR) index was detected to be 0.22. The tetL and aph(3')-IIIa were the most commonly found antibiotic resistance genes in the strains. However, E. mundtii strains HC56.3, HC73.1, HC147.1, and E. faecium strain HC121.4 were detected to lack any of the antibiotic resistance genes examined in this study. Only E. mundtii HC166.3 showed hemolytic activity, while none of the strains engage in gelatinase activity. The strains were identified to have virulence factor genes with a low rate. None of the virulence factor genes could be detected in E. mundtii HC26.1, HC56.3, HC73.1, HC165.3, HC166.8, and E. faecium HC121.4. The E. mundtii HC73.2 strain displayed the highest presence of virulence factor genes, namely gelE, efaAfs, cpd, and ccf. Similarly, the E. mundtii HC112.1 strain showed a significant presence of genes efaAfm, ccf, and acm. There was no decarboxylation of histidine, ornithine, or lysine seen in any of the strains. Nevertheless, E. faecium HC121.4 and HC161.1 strains could decarboxylate tyrosine, but E. mundtii HC26.1, HC56.3, HC73.1, HC73.2, HC112.1, HC147.1, HC155.2, HC165.3, HC166.3, HC166.5, and HC166.8 strains only showed a limited capacity for tyrosine decarboxylation. None of the strains possessed the hdc, odc, or ldc genes, but all of them had the tdc gene.ConclusionThe E. mundtii HC56.3 and HC73.1 strains were deemed appropriate for utilization in food production. Using the remaining 11 strains as live cultures in food production activities could pose a possible risk to consumer health.

Pilot scale production of high-content mycoprotein using Rhizopus microsporus var. oligosporus by submerged fermentation and agro-industrial by-products

While mycoprotein has gained traction as a human food source, its potential as a nutrient for animals remains largely unexplored. The mycoprotein-producing Rhizopus microsporus var. oligosporus, a fungus traditionally used for human food in Indonesia, is promising. It could revolutionise animal nutrition once it is Generally Recognized as Safe (GRAS) and is a biosafety level 1 (BSL1) organism. To enhance sustainably, we propose using sugar cane molasses (SM) and corn steep liquor (CSL) as nutrient sources. Also, we investigated the growth of R. microsporus var. oligosporus in five 14 L external-loop airlift bioreactors using CSL as the sole nutrient source. After 96 h of fermentation, at 25 °C and 0.5 vvm, the mycelium produced had an average biomass yield of 38.34 g L−1, with 70.18 % (m v−1) crude protein (mycoprotein). This bioprocess, which is scalable and economically viable, produces high amounts of mycoprotein for animal feed using CSL, a cost-effective agro-industrial by-product, providing a practical solution to the growing demand for animal protein.

NaCl inhibits Geotrichum citri-aurantii growth by inducing lipid droplets accumulation and enhances the biocontrol efficacy of Kloeckera apiculata 34-9

Decays resulting from infections by Geotrichum citri-aurantii cause a significant reduction in citrus postharvest quality and marketable yield. In the present work, 0.8 mol · L-1 NaCl of Generally Recognized as Safe (GRAS) salts were identified to have complete inhibition activity to G. citri-aurantii in vitro and in vivo. An inhibitory mechanism study showed that 0.1 mol · L-1 NaCl changed the expression of six genes (Gci002087, Gci003354, Gci000013, Gci000394, Gci003505 and Gci004433) in target of rapamycin (TOR) pathway, thus inducing a large number of lipid droplets (LDs) accumulation with TEM observations and staining. In vivo, 0.1 mol · L-1 NaCl could raise up the activities of CAT, PAL, PPO and cellulase which hence to strengthen the ability of resistance tolerance of citrus. In addition, NaCl not only accelerated the growth of the biocontrol yeast Kloeckera apiculata strain 34-9, but also increased the expression level of adhesin-encoding genes and biofilm-formation encoding gene (34-9 - 3691). Finally, the storage experiments in 3 months demonstrated that the combination of NaCl and the yeast 34-9 could keep fresh, reduce decay and prolong its shelf-life, thus implying that the combination of biocontrol strain and GRAS salts has been explored as a promising alternative to synthetic fungicides.

Effects of Combined Cold Plasma and Organic Acid‐Based Sanitizer Treatments Against Salmonella enterica on Tomato Surfaces

ABSTRACTIncidence of foodborne illness due to bacterial contamination of fresh produce continue to exist despite continuous research on processing interventions to mitigate the problem. In this study, we combined atmospheric cold plasma treatments with an antimicrobial solution containing specific organic acids generally recognized as safe (GRAS) by the FDA and tested its antimicrobial efficacy against Salmonella enterica inoculated on tomato surfaces. Tomato surfaces were inoculated with at 5.6 log CFU/g of Salmonella by spotting 0.1 mL of 7 log CFU/ml Salmonella onto the tomato stem scars, and by dipping whole tomatoes into a solution of 7 log CFU/ml Salmonella for 3 min to achieve 4.1 log CFU/g. Antimicrobial efficacy of the organic acid‐based sanitizer + cold plasma treatments for 30, 60, 120, 180, and 360 s, were investigated, and significant bacterial inactivation was achieved above 120 s treatments. At 120 s, surviving populations of aerobic mesophilic bacteria recovered on the tomatoes surfaces averaged < 2 logs/g while yeast and mold survival averaged < 1 CFU/g. Treatment combination with this organic acid‐based sanitizer + cold plasma for 120 s resulted in a 4.9 log reduction of Salmonella on the stem scar area and a 3.9 log reduction on the smooth peel surface. Similarly, populations of aerobic mesophilic bacteria recovered on treated tomato surfaces averaged < 0.3 log CFU/g. The results of this study indicate that combining an organic acid‐based sanitizer with cold plasma treatments for ≥ 120 s can inactivates significant populations of Salmonella to enhance the microbial safety of tomato surfaces designated for fresh‐cut salad.

Antibiotic-resistant mutants of lactic acid bacteria: potential food control agents

Background: One of the most pressing challenges facing global agriculture today is the restoration of degraded farmland. Organic fertilisers are increasingly being used as a safer alternative to synthetic forms. The most promising form of environmentally friendly fertiliser is a bioconsortium based on different groups of microorganisms. Lactic acid bacteria (LAB) with high antifungal activity, isolated from Armenian dairy products, can form part of such a bioconsortium and protect plants from phytopathogens. In addition, LAB can be used as a preservative in the food industry due to its GRAS (generally recognised as safe) status. Objective: To obtain antibiotic-resistant pleiotropic lactic acid bacterial mutants with higher antifungal activity and broad-spectrum activity compared to natural strains. Methods: The objects of the study were LAB, whose antifungal activity was studied against phytopathogenic fungi of the genera Penicillium, Aspergillus, Alternaria and Fusarium, which cause crop/food spoilage. To increase antifungal activity, spontaneous pleiotropic mutants resistant to the streptomycin, kanamycin or rifampicin were obtained on the basis of original strains. Results: Among all the LAB tested, Lactobacillus bovis MDC 1061, L. paracasei MDC 10898 and L. buchneri BKM 1599 were selected for their antifungal activity. Based on the latter, 78 antibiotic-resistant mutants with different levels of antifungal activity were obtained. L. bovis MDC 1061 STR-25 and L. buchneri BKM 1599 RIF-7 mutants with up to 98% and 95% inhibition of fungal growth, respectively, were selected as potential food control agents. Conclusion: Antibiotic-resistant mutants L. bovis MDC 1061 STR-25 and L. buchneri BKM 1599 RIF-7 with high antifungal ability and a wide range of its effects were obtained. For the first time, the phenomenon of pleiotropic effect has been used in LAB to improve antifungal activity. After extensive study, these mutants can be used as promising components for creating an effective bioconsortium, as well as a preservative in food production, especially in the case of plant-based functional foods to protect against contamination by fungi. Keywords: lactic acid bacteria, pleiotropic mutants, antifungal activity

Investigating the Use of Natural Hydrophobic Deep Eutectic Solvents for Extracting Carotenoids from Food and Food By-products

Abstract In today’s food industry, the demand for sustainable methodologies in producing health-promoting natural products is rising. Traditional extraction techniques are increasingly deemed unreliable due to environmental concerns. To address these challenges, the scientific community has developed Green Chemistry , a specialized branch that aims to minimize or eliminate hazardous processes and substances in industrial applications. One promising and eco-friendly alternative emerging from Green Chemistry is the use of NaDESs which are prepared with natural components, generally recognized as safe (GRAS), and offer an environmentally benign option for various industrial processes. They are characterized by their biodegradability, low toxicity, and ability to be tailored for specific applications, making them an ideal choice for green extraction methods. In this context, NaHDESs have shown significant potential as extracting media for bioactive compounds, particularly carotenoids, from food and food by-products. Carotenoids, known for their antioxidant properties and health benefits, are highly sought after in the food and nutraceutical industries. Traditional extraction methods often rely on organic solvents, which pose environmental and health risks. In contrast, NaHDESs offer a safer and more sustainable alternative. This study explores the definition, properties, and applications of NaHDESs, with a particular focus on their use in extracting carotenoids. By examining various food and vegetable by-products, the research aims to evaluate the solvent’s efficiency and effectiveness in this application. The findings could pave the way for eco-friendly extraction techniques in the food industry, aligning with the principles of Green Chemistry and addressing the growing demand for natural and health-promoting products. Information © The Author 2024

High-Yield Expressed Human Ferritin Heavy-Chain Nanoparticles in K. marxianus for Functional Food Development.

The use of Generally Recognized as Safe (GRAS)-grade microbial cell factories to produce recombinant protein-based nutritional products is a promising trend in developing food and health supplements. In this study, GRAS-grade Kluyveromyces marxianus was employed to express recombinant human heavy-chain ferritin (rhFTH), achieving a yield of 11 g/L in a 5 L fermenter, marking the highest yield reported for ferritin nanoparticle proteins to our knowledge. The rhFTH formed 12 nm spherical nanocages capable of ferroxidase activity, which involves converting Fe2+ to Fe3+ for storage. The rhFTH-containing yeast cell lysates promoted cytokine secretion (tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and -1β (IL-1β)) and enhanced locomotion, pharyngeal pumping frequency, egg-laying capacity, and lifespan under heat and oxidative stress in the RAW264.7 mouse cell line and the C. elegans model, respectively, whereas yeast cell lysate alone had no such effects. These findings suggest that rhFTH boosts immunity, holding promise for developing ferritin-based food and nutritional products and suggesting its adjuvant potential for clinical applications of ferritin-based nanomedicine. The high-yield production of ferritin nanoparticles in K. marxianus offers a valuable source of ferritin for the development of ferritin-based products.

435 Use of plant bioactives to improve the efficiency of rumen function

Abstract Plant bioactives, also known as phytogenics or phytobiotics, are secondary metabolites of plant origin that are deployed as host-defense mechanisms against predators or to enable them to be more competitive in their environment. These metabolites also regulate growth and reproductive functions in the plant. They can be grouped into three major classes: terpenoids, phenolic compounds, and alkaloids. Various studies have shown their worth in improving livestock production and health via insectifugal, organoleptic and enhanced rumen functions. Ruminants are critical to food security because they are able to convert poor-quality feed materials into nutritious products like meat. The rumen provides temporary storage of feed but serves the main function of digesting those complex materials through symbiotic relationships among prokaryotic and eukaryotic organisms within the rumen vat. Antimicrobial resistance and other health or social license challenges have led researchers to find acceptable replacements that will not exacerbate the health and social issues in animals and humans. Plant bioactives appeal to producers because of the ‘natural’ brand and because they are usually certified as generally recognized as safe (GRAS). Anecdotal reports from input companies show an increased demand for mineral supplements infused with phytogenics. The complex chemical compositions of these products appear to make them less prone to developing resistance by microorganisms. Phytogenics research has been further catalyzed by government legislation that has banned the use of medically important antibiotics to prevent the spread of antibiotic resistance. Potential improvements in livestock productivity are associated with mechanisms such as enhanced nutrient utilization, increased digestibility, modified digestive secretions or altered microbial communities. These benefits contribute to the increasing use of these phytogenics by livestock producers to keep their animals healthy and productive while enhancing the profitability of their operations. We will examine the different bioactives and their applications in improving the efficiency of the rumen function. Some commercial products in the Canadian market will be presented. Other research trials showcasing their potential benefits to rumen health and general animal productivity will also be discussed.

Recovery of Phenolic Compounds by Deep Eutectic Solvents in Orange By-Products and Spent Coffee Grounds

Orange and coffee grounds by-products, rich in phenolic bioactive compounds, can be used in the food industry as antioxidants, colorants, flavorings and additives, mainly because they are solvents that are easy to prepare, have a lower cost, are thermally stable, biodegradable, renewable, and are considered GRAS (Generally Recognized as Safe). Deep eutectic solvents, which are sustainable and have lower melting points, are effective for extracting these compounds. This study aimed to evaluate the use of deep eutectic solvents (DES) in extracting Total Phenolic Compounds (TPC), from orange by-products and spent coffee grounds. DES formed by citric acid: mannitol (CM-DES), and lactic acid: glucose (LG-DES), were evaluated by varying the following parameters: water content (10–50%), solid–liquid ratio (1:5–1:50 w/w) and temperature (40–50 °C). DES citric acid: mannitol presented the best efficiency in the extraction of TPC under the conditions of 10% water, 80 °C, and solid–liquid ratio 1:10 (w/w) for the orange by-products (1782.92 ± 4.50 mg GAE/L) and 1:15 (w/w) for spent coffee grounds (1620.71 ± 3.72 mg GAE/L). The highest antioxidant activity was observed in the extraction with CM-DES for both by-products in the three methods evaluated: Ferric Reducing Antioxidant Power (FRAP) (1.087 ± 0.004 and 1.071 ± 0.006 mol ascorbic acid/L), DPPH radical scavenging activity (2,2-difenil-1-picrilhidrazil—DPPH) (0.233 ± 0.003 and 0.234 ± 0.001 mol Trolox equivalent/L), and radical cation scavenging activity ABTS (2,2-azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid—ABTS) (0.284 ± 7.16 and 0.319 ± 0.002 mol Trolox equivalent/L). Therefore, DES with citric acid: mannitol is a promising alternative to conventional solvents to recover phenolic compounds in agro-industrial by-products, such as orange by-products and SCG.

Antifungal and Antioxidant Properties of Some Artificial Antioxidants, Generally Recognized as Safe Compounds and Nano-Oxides

In this study, the usage potential of some artificial antioxidants, generally recognized as safe (GRAS) compounds and nano-oxides solutions in wood preservation industry, was investigated. For this purpose, antifungal and antioxidant properties of solutions were determined. Erythorbic acid, ethoxyquin, potassium disulfide, sodium ascorbate, sodium erythorbate and Enginerring and Nature Sciences Faculty (TBHQ) were selected as artificial antioxidants; dehydroacetic acid, sorbic acid and sodium benzoate were used as GRAS compounds and nano MgO, nano CeO, nano ZnO, nano SiO2 and TiO2 were investigated as nano-oxides in this study. Three different concentrations (0.5%, 1.0% and 1.5% ) were prepared, and anti-fungal test were carried out. The brown rot fungus Coniophora puteana (Schumach.) P. Karst. (BAM Ebw. 15) was used for the anti-fungal test. Then antioxidant activity of the solutions were determined. Iron (III) ion reducing antioxidant power method (FRAP) was used to determine the antioxidant activity of solutions. All solutions at 1.5% concentration completely inhibited the growth of C. puteana fungus. The antioxidant activity of solutions was sorted as artificial antioxidants>GRAS compounds>nano-oxides, respectively. It was concluded that the tested substances can be used as impregnating agents in wood preservation.

Sinapic acid-pullulan based inflammation responsive nanomicelles for the local treatment of experimental inflammatory arthritis

Rheumatoid arthritis (RA) is a chronic inflammatory disorder of joints. It is one of the major causes of disability and morbidity worldwide. Administration of conventional drugs through the systemic route restricts the bioavailability of drugs, systemic toxicity, and reduced efficacy. We have introduced Rebamipide (Reb)-loaded Sinapic acid (SA)-Pullulan (PL) nanomicelles (Reb@SA-PL NMs), a nanotechnology based drug delivery system for the treatment of inflammatory arthritis. PL is a polysaccharide obtained from the fungus Aureobasidium pullulans, and SA is a bioactive polyphenol found in various plants. Both are classified by US-FDA Generally Recognised as Safe (GRAS) materials. Reb@SA-PL NMs found to be cytocompatible. Subsequently, intra-articular administration of Reb@SA-PL NMs enhances the anti-arthritic potential compared to free Reb drug in collagen-induced experimental inflammatory arthritis rat model. Reb@SA-PL NMs reduced the expression of RANKL receptor and Nf-κB. Reb@SA-PL NMs reverses the breakdown of type II collagen, MMP-13, and inhibits the pro-inflammatory markers. Reb@SA-PL NMs prevented bone erosion, cartilage degradation, joint oedema, and synovial inflammation. The results of the study demonstrated that Reb@SA-PL NMs, an enzyme-responsive drug delivery system, has excellent potential for alleviating inflammatory arthritis by blocking MMP-13 and RANKL.

Human milk oligosaccharides produced by synthetic biology

The benefits of human breast milk (HBM) to newborn growth, development, and overall health have been well investigated. As breastfeeding rate has declined significantly, the use of infant formula has risen and remains a significant component of the infant's diet. HBM contains several essential nutritional components, with human milk oligosaccharides (HMO) being the third most abundant following lactose and lipids. A diverse array of HMOs is known to exert great benefits in intestinal, immune, and cognitive functions. In contrast to HBM, infant formula containing mainly bovine milk lacks the diversity of HMOs. Efforts have been made to replicate this characteristic in infant formula through chemical and chemoenzymatic synthesis as well as microbial production. Utilizing microbial hosts appears to be more favourable due to its accessibility and cost-efficiency. Escherichia coli has been preferably used due to its high incorporation of DNA, high-level expression capability, and rapid growth. However, potential endotoxin contamination raises health concerns and prevents approval as Generally Recognized as Safe (GRAS) by the FDA. This prompts the use of other microbes such as Bacillus subtilis. Future research in this area is needed to optimize effective procedures using microbial hosts, high yield production, and economic feasibility. This may lead to infant formulas closely mimicking HBM and its health benefits.

Reprogramming Yarrowia lipolytica metabolism for efficient synthesis of itaconic acid from flask to semipilot scale.

Itaconic acid is an emerging platform chemical with extensive applications. Itaconic acid is currently produced by Aspergillus terreus through biological fermentation. However, A. terreus is a fungal pathogen that needs additional morphology controls, making itaconic acid production on industrial scale problematic. Here, we reprogrammed the Generally Recognized As Safe (GRAS) yeast Yarrowia lipolytica for competitive itaconic acid production. After preventing carbon sink into lipid accumulation, we evaluated itaconic acid production both inside and outside the mitochondria while fine-tuning its biosynthetic pathway. We then mimicked the regulation of nitrogen limitation in nitrogen-replete conditions by down-regulating NAD+-dependent isocitrate dehydrogenase through weak promoters, RNA interference, or CRISPR interference. Ultimately, we optimized fermentation parameters for fed-batch cultivations and produced itaconic acid titers of 130.1 grams per liter in 1-liter bioreactors and 94.8 grams per liter in a 50-liter bioreactor on semipilot scale. Our findings provide effective approaches to harness the GRAS microorganism Y. lipolytica for competitive industrial-scale production of itaconic acid.

Regulation of Added Substances in the Food Supply by the Food and Drug Administration Human Foods Program.

The US food supply is increasingly associated with diet-related diseases, toxicity, cancer, and other health harms. These public health concerns are partly attributable to a loophole in federal law. The Food and Drug Administration (FDA) evaluates the premarket safety of ingredients regulated as food additives but allows the food industry to self-regulate and determine which substances to classify as generally recognized as safe (GRAS) based on undisclosed data and conclusions that the FDA never sees. Furthermore, the FDA lacks a formal approach for reviewing food additives and GRAS substances already found in the food supply. Substances in the food supply thus include innocuous ingredients (e.g., black pepper), those that are harmful at high levels (e.g., salt), those that are of questionable safety (e.g., potassium bromate), and those that are unknown to the FDA and the public. A recent court decision codified these gaps in the FDA's current approach, leaving states to try to fill the regulatory void. The FDA and Congress should consider several policy options to ensure that the FDA is meeting its mission to ensure a safe food supply. (Am J Public Health. Published online ahead of print August 8, 2024:e1-e10. https://doi.org/10.2105/AJPH.2024.307755).

Effects of Probiotics on Liver and Kidney Function Parameters

This study aimed to study the effects of probiotics on liver and kidney function parameters. An in vitro study was used to identify food-grade probiotics for human consumption. As far as probiotic strains used, B. longum subspecies infantis, L. acidophilus, as well as B. bifidum are all GRAS (Generally Recognized as Safe). Over the course of six months, 100 patients with CKD and Liver disease 50 patients were without treatment and 50 patients were given two capsules each morning and evening containing 2.5*109 CFU B. longum subspecies infantis, L. acidophilus, as well as B. bifidum at a ratio of 1:1:1, and 50 normal indivisuals were used in this study. Pre-test nutritionists recommended the participants to eat low-protein diets and to avoid any lifestyle or dietary changes throughout the experiment time. Blood was collected for evaluation of AST, ALT, Total protein, Urea, and creatinine levels in both patients and normal. These kits were obtained from Biolabo company, these were done according to company instructions. The results showed that the probiotics had a therapeutics effect which causes a significant difference in all parameters in treated patients as compared with patients that were not treated. In conclusion, Probiotics have therapeutic effects on the injured kidney and the liver, which are indicated by decreasing the liver and kidney functions.

Optimization of Fermentative Parameters to Improve Gamma-Aminobutyric Acid (GABA) Production by Lactiplantibacillus plantarum B13

Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the mammalian brain, and it possesses several physiological properties, such as depression reduction and anxiety release. GABA production by microbial synthesis is favoured over other methods like enzymatic and chemical synthesis due to the ease of operation and less formation of harmful pollutants. Lactic acid bacteria are widely applied for GABA production because of high GABA yield and their Generally Recognized as Safe (GRAS) status , which is critical in food and ingredient safety. In this study, various fermentation conditions, including incubation time, pH, temperature, monosodium glutamate (MSG) concentration, pyridoxal-5’-phosphate (PLP) concentration and glucose concentration were screened by one-factor-at-a-time strategy to achieve the optimal GABA production by a potential probiotic strain, Lactiplantibacillus plantarum B13. The result revealed the strain exhibited the optimal GABA production of 19.073 ± 0.5214 g/L with the highest GABA productivity of 0.424 g/L/h under fixed conditions: incubation time of 66 hours, pH 5.5, temperature of 35°C, MSG concentration of 5% (w/v), PLP concentration of 0.7 mM PLP and glucose concentration of 60 g/L. The findings of this study show that fermentation parameters are dependent on species and strains due to the different properties of glutamic acid decarboxylase enzymes and optimization of single parameters is important as a preliminary step to identify the range of fermentation factors that affect GABA yield prior further research endeavours. This study also has great implications for GABA production by L. plantarum B13 and provides a prerequisite for developing new healthy products enriched with GABA as daily supplements to support relaxation and regulate mood, reduce stress and promote better sleep.

Bacteriophages: Natural antimicrobial bioadditives for food preservation in active packaging

Developing innovative films and coatings is paramount for extending the shelf life of numerous food products and augmenting the barrier and antimicrobial properties of food packaging materials. Many synthetic chemicals used in active packaging and food storage have the potential to leach into food, posing long-term health risks. It is imperative for active packaging materials to inherently possess biological protective properties to ensure food quality and safety throughout its storage. Bacteriophages, or simply phages, are bacteria-eating viruses that serve as promising natural biocontrol agents and antimicrobial bioadditives in food packaging materials, specifically targeting bacterial foodborne pathogens. These phages are generally recognized as safe (GRAS) by regulatory authorities for food safety applications. They exhibit targeted action against various Gram-positive and -negative foodborne pathogens, including Bacillus spp., Campylobacter spp., Escherichia coli, Listeria monocytogenes, Salmonella spp., Shigella spp., and Vibrio spp., associated with foodborne spoilage and illness without affecting the beneficial microbes. Phage cocktails can be applied directly on food surfaces, incorporated into food packaging materials, or utilized during food processing treatments. Unlike chemical agents, phage activity increases proportionally with the rise in pathogenic bacterial populations. Researchers are exploring various packaging materials to deliver phages with broad host range, stability, and viability ensuring their effectiveness in safeguarding various food systems. The effectiveness of phage immobilization or encapsulation on active food packaging materials depends on various factors, including the characteristics of polymers, the choice of solvents, the type of phage, and its loading efficiency. Factors such as the orientation of phage immobilization on substrates, pH, temperature, exposure to carbohydrates and amino acids, exopolysaccharides, lipopolysaccharides, and metals can also influence phage activity. In this review, we comprehensively discuss the various active packaging systems utilizing bacteriophages as natural biocontrols and antimicrobial bioadditives to reduce the incidence of foodborne illness and enhance consumer confidence in the safety of food products.

The Frequently Used Industrial Food Process Additive, Microbial Transglutaminase: Boon or Bane.

Microbial transglutaminase (mTG) is a frequently consumed processed food additive, and use of its cross-linked complexes is expanding rapidly. It was designated as a processing aid and was granted the generally recognized as safe (GRAS) classification decades ago, thus avoiding thorough assessment according to current criteria of toxicity and public health safety. In contrast to the manufacturer's declarations and claims, mTG and/or its transamidated complexes are proinflammatory, immunogenic, allergenic, pathogenic, and potentially toxic, hence raising concerns for public health. Being a member of the transglutaminase family and functionally imitating the tissue transglutaminase, mTG was recently identified as a potential inducer of celiac disease. Microbial transglutaminase and its docked complexes have numerous detrimental effects. Those harmful aspects are denied by the manufacturers, who claim the enzyme is deactivated when heated or by gastric acidity, and that its covalently linked isopeptide bonds are safe. The present narrative review describes the potential side effects of mTG, highlighting its thermostability and activity over a broad pH range, thus, challenging the manufacturers' and distributers' safety claims. The national food regulatory authorities and the scientific community are urged to reevaluate mTG's GRAS status, prioritizing public health protection against the possible risks associated with this enzyme and its health-damaging consequences.