Postbiotics at the interface of microbial biotechnology and therapeutics: industrial production, functional mechanisms, and clinical potentials.

Probiotical cell fragments (PCFs) are structural components of the probiotic cell lysate(s) and exhibit similar beneficial effects on the host as live probiotic bacteria. With cell fragment technology (CFT?), the structural fragments are isolated and purified from live probiotic cells. While observed to be strain-dependent as in the case of live probiotics, orally administered PCFs demonstrated a broad spectrum of immune modulation functions; anti-allergy; anti-inflammation; anti-bacterial and anti-viral properties; anti-mutagenic; and radioprotective and detoxification abilities in humans and animals. The PCFs mechanisms of action include events of motifs of cell wall peptidoglycans, DNA motifs, nucleotide containing components, lipoteichoic acids (LPAs), surface layer (S-layer) proteins, and cytoplasmic proteins. Different immunological in vivo-in vitro tests have shown that PCFs, essentially, have the ability to stimulate the macrophages, and induce cytokines such as interleukins, tumor necrosis factors (TNFs), interferons (IFNs), and natural killer (NK) cells. PCFs may be used as ingredients for foods and beverages or as nutritional supplements with long term stability and shelf-life up to 5 years. PCFs may also be used as health restorative ingredients in cosmetic products. The outcome of probiotics CFT? stands as an advantage to the food and pharmaceutical industries, regarding the formulation of novel products with unadulterated sensory characteristics of origin. Hence, PCFs are being characterized here as “novel nutraceutical ingredients” for health maintenance in both humans and animals.

The main input of energy and matter (excluding water and oxygen) in aquaculture systems are fertilized eggs, food, and fertilizer, while waste and harvest represent the main output. Aquaculture systems are classified by stocking density, yield, space efficiency, and the extent of reliance on ecosystem services or technology. Ecologically sustainable aquaculture is promoted by collection, processing, and proper disposal of waste and by trophic recycling of waste within the system. Intensive recirculating aquaculture systems (RAS) and extensive aquaculture are most ecologically sustainable. Semi-intensive, open, cage, raceway, and net pen aquaculture systems are least ecologically sustainable. Aquaculture sustainability is increased when feed efficiency is maximized, and feed conversion rate minimized by optimal composition, consistency, and application of food. Live food is critical in hatcheries for feeding larvae. It consists of phytoplankton (microalgae) and zooplankton (rotifers, brine shrimp, and copepods). Formulated aquaculture feeds containing optimal amounts and ratios of amino acids, proteins, lipids, carbohydrates, minerals, vitamins, and other essential nutrients have been developed. An optimally nutrient-balanced diet based on formulated feeds maximizes the health of aquaculture animals. Aquaculture waste is produced from unused decaying food, excreted animal waste, and decaying plant, bacterial, and animal biomass. Aquaculture waste is treated by mechanical filtration and sedimentation, biological filtration, chemical filtration, degassing, oxygenation, and sterilization. Stress induced by poor water quality impairs the welfare and performance of aquaculture organisms. Universal, non-specific symptoms of stress are routinely monitored in aquaculture. They include animal behaviour and appearance, haematocrit, lymphocyte counts, and plasma cortisol and glucose concentrations.

Bioactive metabolites are a substance that has a biological activity and should include all microbial compounds obtained either from microbes or from any other living thing. Around 23,000 bioactive metabolites produced from microorganisms, out of which 10,000 (45%) are produced by Actinobacteria alone. Actinomycetes are known to produce an extensive range of bioactive metabolites as well as variety of enzymes with multiple biotechnological applications and important for pharmaceutical, food, agricultural, and environmental applications. Bioactive metabolites are widely used and studied for obtaining antibiotics, antifungals, antivirals, immunosuppressants, compounds with anticancer and antioxidant activity, enzymes, bioinsecticides, biostimulants, biosurfactants and other applications. Bioactive metabolites derived from Actinomycetes are more attractive than bioactive metabolites from other sources because of their high stability and unusual activity specificity. The main applications of Actinomycetes, studies have focused on antimicrobial potential, enzymes production, agricultural uses, bioremediation, and others related to their secondary metabolites production. The review aimed to summarize information about the potentials of Actinomycetes novel bioactive metabolites with their applications in different prospects.

For a long time, probiotics have been widely used as safe microorganisms that can confers a health benefit effects on the host, directly or indirectly. Recently, postbiotics have gained interest as new health promoters. Postbiotics have recently been defined as complex mixture of functional bioactive compounds secreted by probiotics during a fermentation process (such as biosurfactants, proteins, short chain fatty acids, organic acids, bacteriocins, vitamins etc.). According to current data, postbiotics have advantages over live probiotics with regard to: ease extraction, standardization, and storage, availability for industrial-scale-up, specific mechanism of action, impossible to transfer and acquire antibiotic resistance genes and their interaction with the cellular receptors to trigger the targeted responses. However, several aspects related to postbiotics have not been fully elucidated. Here, we provided a critical review of the postbiotic definition, mechanisms of action, underlying their beneficial effects, as well as current trends for applications in foods and pharmaceuticals.

The benefits of probiotics in foods are well-studied, but maintaining their effectiveness requires survival through various stress conditions like processing, storage, and digestion. Safety concerns, such as antibiotic resistance and potential infections, have raised questions about using live probiotics. Although traditionally, probiotic foods require live microorganisms for beneficial effects, research suggests that inactivated (non-viable) probiotic cells and their metabolic byproducts can also provide health benefits. This has led to the introduction of terms like postbiotics (cell-free metabolic byproducts of probiotics) and parabiotics (inactivated probiotic cells and cell compounds). Most studies focus on Lactic Acid Bacteria (LAB) as sources of postbiotics and parabiotics, but there is limited research on yeast-derived compounds. Yeasts can produce diverse metabolites and unique intracellular components, potentially enhancing shelf-life, sensory qualities, safety, and health benefits in food products. This review explores the advances in bacterial and yeast postbiotics and parabiotics and their applications in food, pharmaceuticals, and feed systems.

Total and viable microbial cell counts are increasingly important for applications including live biotherapeutic products, food safety, and probiotics. In microbiology, cells are quantified using methods such as colony forming unit (CFU), flow cytometry, and polymerase chain reaction (PCR), but different methods measure different aspects of the cells (measurands), and results may not be directly comparable across methods. In the absence of a ground-truth reference material for cell count, one cannot quantify the accuracy of any cell counting method, which limits method performance assessments and comparisons. Herein, a modified analysis of cell counting methods based on the ISO 20391-2:2019 standard was developed and demonstrated for microbial cell samples diluted over a log-scale range of concentrations. Escherichia coli samples ranging in concentration from ~5 x 105 cells/mL to 2 x 107 cells/mL were quantified using CFU, Coulter principle, fluorescence flow cytometry, and impedance flow cytometry. Quality metrics modified from the ISO standard were calculated for each method and shown to be repeatable across replicate experiments. The quality metrics illustrate large differences in proportionality and variability across methods, with total cell counts in good agreement and viable cell count having more variability. As the ISO standard is meant to guide fit-for-purpose method selection, interpretation of the results and quality metrics can drive method choice and optimization. The framework introduced here will help researchers select fit-for-purpose counting methods for quantification of microbial total and viable cells across a range of applications.

Graphical abstract Abstract 7-Ketocholesterol (or 7-oxocholesterol) is an oxysterol essentially formed by cholesterol autoxidation. It is often found at enhanced levels in the body fluids and/or target tissues of patients with age-related diseases (cardiovascular, neuronal, and ocular diseases) as well as in subjects concerned with civilization diseases (type 2 diabetes, bowel diseases, and metabolic syndrome). The involvement of increased 7-ketocholesterol levels in the pathophysiology of these diseases is widely suspected. Indeed, 7-ketocholesterol at elevated concentrations is a powerful inducer of oxidative stress, inflammation, and cellular degeneration which are common features of all these diseases. It is important to better know the origin of 7-ketocholesterol (diet, incidence of environmental factors, and endogenous formation (autoxidation and enzymatic synthesis)) and its inactivation mechanisms which include esterification, sulfation, oxidation, and reduction. This knowledge will make it possible to act at different levels to regulate 7-ketocholesterol level and counteract its toxicity in order to limit the incidence of diseases associated with this oxysterol. These different points as well as food and biomedical applications are addressed in this review.

Specialized Bioactive Microbial Metabolites: From Gene to Product.

Paraprobiotics: Evidences on their ability to modify biological responses, inactivation methods and perspectives on their application in foods

Pearl millet is an arid region crop grown under adverse agro-climatic conditions. It has high energy value (361 Kcal/100 g) compared to other cereals (325–349 Kcal/100 g). Its carbohydrate content is 67.5 g/100 g with high fiber and α amylase activity. It is a gluten free grain, retains its alkaline properties after cooking, and is suitable for persons suffering from gluten intolerance. It has quality protein and the highest amount of fat content rich in unsaturated fatty acids (75%). It is a well-known source of B-complex and lipid soluble vitamins. This millet is the richest source of microminerals, particularly iron, phosphorous and zinc. Micronutrient deficiencies are responsible for major health problems among the poor in developing nations. Owing to its good nutritional profile, it has numerous health benefits. It is an underutilized millet; therefore, it is imperative to spread the knowledge and importance of millet’s nutritional profile and reorient the effort to generate demand through value addition and quality improvement. Biofortification is an attractive means of increasing the concentration of plant-derived nutrients in the edible organ, over and above the normal, through breeding new varieties. It is an economically feasible, easily implementable technique of delivering micronutrients to poor sections of population suffering micronutrient malnutrition. The biofortification of pearl millet is under way in India, and seven biofortified hybrids/varieties enriched with a higher amount of iron and zinc have been developed. Despite being nutrient rich, its food applications are limited due to rancidity, which reduces its shelf stability. Pearl millet’s high polyunsaturated fatty acid profile, along with the presence of lipases, is what causes rancidity and off-flavors. Screening of low rancidity susceptible pearl millet genotypes and understanding and modifying the enzymatic pathways may help to overcome this problem. Thus, development of various kinds of value added products from biofortified hybrids/varieties and making people aware of the importance and benefits of their consumption will help in addressing the malnutrition problem of growing populations.

Effects of bacterial inactivation methods on downstream proteomic analysis

The Northwestern University Clinical and Translational Sciences Institute (NUCATS) was launched in 2007 to create a central hub supporting clinical and translational science (CTS) across numerous schools at Northwestern University, our three main clinical partners (Northwestern Memorial Healthcare Corporation, Ann and Robert H. Lurie Children's Hospital, and The Rehabilitation Institute of Chicago), community and industry stakeholders, and beyond. NUCATS is designed to support the entire spectrum of CTS, from basic discovery through clinical trials to community-based research, dissemination and implementation, regardless of disease area. Funded by a Clinical and Translational Award (CTSA) from the NIH, and supported by extensive institutional resources, NUCATS has become an innovation leader in several key areas of CTS. Here we highlight the critical contributions an enterprise data warehouse can make to advancing translational science and quality clinical care to enable a learning healthcare system. Central to the success of our EDW has been the governance model and data structure that have enabled rapid advances in CTS, several examples of which we provide. Informatics platforms that enable CTS have been a major focus of activity in NUCATS. The Northwestern University Biomedical Informatics Center was created to bring together informatics activities across NUCATS' partners. In 2015, the role of the center was expanded to explicitly include big data/data science, and the name of the center was changed to the Center for Data Science and Informatics (CDSI). CDSI brings together biomedical informatics researchers and clinical informatics leaders from NUCATS' partners into an organization to coordinate biomedical informatics across the NU academic medical enterprise. To meet this mission, CDSI has culled the necessary expertise and resources to enable and facilitate the application of informatics solutions to clinical and translational research. CDSI-coordinated infrastructure is a crucial component of translational research at Northwestern. A central component of the informatics infrastructure of CDSI is the Northwestern Medicine Enterprise Data Warehouse (NMEDW). Created in 2007 as part of the original NUCATS formulation, the NMEDW serves as the primary vehicle for data integration and transfer for both research and clinical operations. The NMEDW was created with an initial $4.6 million, 3-year investment shared among the Feinberg School of Medicine (FSM), the Northwestern Medical Faculty Foundation and Northwestern Memorial Hospital; that investment has grown to $18 million over 8 years. The latter two members have since merged, creating NMHC. From the beginning, the NMEDW was designed to serve both research and clinical needs from a single, unified warehouse. This dual-use model is one of the major strengths of the NMEDW, and one that enables it to function as a unique bridge, integrating healthcare and research, as well as ensuring support from both the research and clinical partners. The NMEDW currently stores over 67 billion observations on 2.9 million unique patients. Each night it loads 44 million new data elements from 76 separate sources including electronic health records (EHR), pathology data from the hospital and research laboratories, biomarker data from research databases, and research transactional data from our eIRB and other institutional systems. It then transforms source data into integrated versions providing access to biological data along with patient demographics and clinical observations, outcomes, and clinical trials protocols. The NMEDW uses the clinical-grade network, computational, and security infrastructure of NMHC to guarantee data security, and is security-audited every year. Use of the NMEDW continues to grow rapidly, showing 261% growth since 2011 (see Figure 1). Bringing together research and clinical data has been critical to the success of phenotyping in the Electronic Medical Records and Genomics (eMERGE) project.1 The NMEDW is governed by a nine member steering committee representing both clinical and research users, and chaired by the NUCATS informatics lead. Annual major project prioritization is conducted using a Delphi process by the steering committee. Each member organization submits a number of proposed projects, NMEDW staff then estimates approximate effort required for each, and the steering committee members create a composite priority ranking. In addition to the steering committee, an NMEDW Advisory Committee, consisting of the CIOs of the member organizations and the NUCATS informatics lead, meets biweekly. The NMEDW is central to clinical operations, supporting, for example, Meaningful Use, outcomes, quality, compliance and revenue cycle reporting. In fact, the NMEDW was the first warehouse to achieve certification for both Meaningful Use Stage 1 and Stage 2 reporting. The NMEDW also serves the role of an "Honest Broker"2 for clinical information to the research community. Access to NMEDW data requires approval from the appropriate oversight body. For quality improvement initiatives, projects must be approved by the respective institutions' quality improvement committee. For research projects, IRB approval is required. The NMEDW also supports an i2b2 instance for feasibility counts without IRB approval. To facilitate the rapid review and approval of data requests, the NMEDW has developed an efficient data steward model, supported by electronic workflow. Each member institution has a designated data steward who reviews all data requests. For access to research data sets, the project Principle Investigator serves as data steward. To support the research needs of individual investigators, the NMEDW staff creates "sandbox" data marts. These data marts contain IRB-approved or quality-committee-approved data elements so researchers are empowered with the data they need without the burden or risk of having data that are not required for a particular project. Since 2009, the NMEDW has supported 858 research projects and has proved to be an enabler of numerous clinical quality improvement initiatives by all contributing clinical affiliates. One example of the value of an integrated research/care EDW has been the experience of Dr. Sanjiv Shah. Heart failure with preserved ejection fraction (HFpEF) remains one of the most common, and most vexing, problems faced by clinicians today. Dr. Shah joined NU in 2007 and initiated the NHLBI-funded TOPCAT study (spironolactone vs. placebo for patients with HFpEF) as site principal investigator in 2008. From the outset, he worked with NUCATS on his recruitment strategy. We designed a customized daily NMEDW-generated report of patients hospitalized at Northwestern Memorial Hospital who fit prespecified criteria based on free text, laboratory, imaging, and medication data. Despite starting enrollment 2 years after the trial began, Northwestern was the top US enrolling site (among 233 total sites in 6 countries), with 77 participants (2.2% of total).3 The same approach is now being used in the NEAT–HFpEF study, funded by the NHLBI Heart Failure Clinical Research Network. Again, Northwestern is the top enrolling site to date. As important as it was to support Dr. Shah's research, the NMEDW-enabled research is also changing care at NMHC. Dr. Shah's clinical HFpEF Program was the first of its kind and uses the same NMEDW-based strategy to identify patients who would benefit from follow up in the HFpEF Clinic. Using the NMEDW, he recently published the first study to conduct high-density phenotypic classification ("pheno-mapping") of HFpEF, defining three discrete, clinically relevant phenogroups of HFpEF patients with significant differences in etiology/pathophysiology and risk for adverse outcomes. Compared with patients in Group 1, those in Groups 2 and 3 are at 3.0- and 4.4-fold higher risk, respectively, for hospitalization or death.4 This discovery is driving research into changing the therapeutic approach to HFpEF. Dr. Emilie Powell wanted to evaluate quality metrics for sepsis care in the emergency department. Sepsis patients are not well identified by diagnostic codes alone, making identification of patients difficult and labor-intensive to process manually. Working with the NMEDW team, Dr. Powell developed an algorithm combining diagnosis codes with clinical parameters to identify 376 severe and/or septic shock patients from 2006 to 2008. This information was used to develop typical patient cases that were the basis for an in situ simulation of sepsis treatment in the Northwestern Memorial Hospital ED, and educational simulations for emergency medicine residents-in-training. The NMEDW sepsis analysis was extended into ongoing projects for Master in Public Health students, medical students and research fellows and has resulted in two publications to date.5, 6 Managing transplant quality is challenging, as CMS transplant scorecards lag current data by up to two years. To help solve this problem, Dr. Bing Ho, transplant nephrologist at NMHC, enlisted the NMEDW to develop a dashboard to analyze kidney transplant data at NMHC. The Real-time Analytics & Process Improvement Dashboard (RAPID) was released in January, 2013. RAPID pulls information from the NMEDW relevant to patient outcomes and quality metrics, replacing a slow manual process. Dr. Ho and his team used RAPID for a root-cause analysis that led to an improvement in patient and allograft survival. The American Society of Transplant Surgeons now encourages the use of RAPID by all member transplant centers. As of mid-2014, 53 transplant centers had registered and downloaded RAPID. Enterprise Data Warehouses that simultaneously support both research and clinical care as joint primary missions, as opposed to warehouses tailored primarily for research or care, are not only viable, but provide significant advantages. These advantages include economies of scale, as well as rapid translation of research discoveries back into clinical practice. However, our successful experience to date suggests that attention to governance models and data stewardship models are critical to ensure stability and efficiency.

From the preparation of bread, cheese, beer, and condiments to vegetarian meat products, fungi play a leading role in the food fermentation industry. With the shortage of global protein resources and the decrease in cultivated land, fungal protein has received much attention for its sustainability. Fungi are high in protein, rich in amino acids, low in fat, and almost cholesterol-free. These properties mean they could be used as a promising supplement for animal and plant proteins. The selection of strains and the fermentation process dominate the flavor and quality of fungal-protein-based products. In terms of function, fungal proteins exhibit better digestive properties, can regulate blood lipid and cholesterol levels, improve immunity, and promote gut health. However, consumer acceptance of fungal proteins is low due to their flavor and safety. Thus, this review puts forward prospects in terms of these issues.

Curcumin, the principal bioactive compound in turmeric (Curcuma longa L.), is widely recognized for its pharmacological properties, including antioxidant, anti-inflammatory, and anticancer activities. However, its low bioavailability remains a major obstacle in the development of curcumin-based applications in food and pharmaceutical sectors. This review provides a comprehensive overview of recent technological advancements aimed at enhancing curcumin’s bioavailability, including encapsulation techniques, lipid-based delivery systems, and chemically modified curcumin derivatives. These innovations have demonstrated significant potential in improving the solubility, stability, and absorption of curcumin in the human body. Furthermore, recent trends in research utilizing natural carriers such as plant-derived proteins and polysaccharides are discussed, aligning with sustainable and food-safe delivery approaches. The review emphasizes an interdisciplinary approach that integrates food material science, biodegradable packaging, bioactive compound chemistry, and nanotechnology engineering. As formulation technologies continue to evolve, the application of curcumin in functional foods and health supplements becomes increasingly promising. The article also highlights existing research gaps and future directions, focusing on biological efficacy, long-term safety, and production scalability. This review aims to serve as a valuable reference for researchers and industry stakeholders in accelerating the utilization of curcumin through effective and sustainable smart delivery systems.

Recent advances of recycling proteins from seafood by-products: Industrial applications, challenges, and breakthroughs