Rhizosphere bacterial exopolysaccharides: composition, biosynthesis, and their potential applications.

Bacterial exopolysaccharides (EPS) are biopolymers of carbohydrates, often released from cells into the extracellular environment. Due to their distinctive physicochemical properties, biocompatibility, biodegradability, and non-toxicity, EPS finds applications in various industrial sectors. However, the need for alternative EPS has grown over the past few decades as lactic acid bacteria's (LAB) low-yield EPS is unable to meet the demand. In this case, rhizosphere bacteria with the diverse communities in soil leading to variations in composition and structure, are recognized as a potential source of EPS applicable in various industries. In addition, media components and cultivation conditions have an impact on EPS production, which ultimately affects the quantity, structure, and biological functions of the EPS. Therefore, scientists are currently working on manipulating bacterial EPS by developing cultures and applying abiotic and biotic stresses, so that better production of exopolysaccharides can be attained. This review highlights the composition, biosynthesis, and effects of environmental factors on EPS production along with the potential applications in different fields of industry. Ultimately, an overview of potential future paths and tactics for improving EPS implementation and commercialization is pointed out.

Efficacy of the incorporation between self-encapsulation and cryoprotectants on improving the freeze-dried survival of probiotic bacteria.

AIMS: This study aimed to improve the viability of probiotic bacteria during freeze-drying by the combination of self-encapsulation and cryoprotectants. METHODS AND RESULTS: Lactiplantibacillus plantarum VAL6 and Lactobacillus acidophilus VAR1 were exposed to environmental stresses including temperature, pH and increased CO2 concentration before performing freeze-drying with the addition of cryoprotectants. The results proved that tested stresses can stimulate the bacteria to synthesize more extracellular polymeric substances to form self-encapsulation that increases their freeze-dried viability. In combination with cryoprotectants to form double-layered microencapsulation, L. plantarum VAL6 stressed at pH 3.5 in combination with whey protein isolate could achieve the highest Improving Cell Viability of 4361-fold, while L. acidophilus VAR1 stressed at 25o C in combination with alginate gave a maximum Improving Cell Viability of 73.33-fold. CONCLUSIONS: The combination of self-encapsulation and cryoprotectants significantly improves the freeze-dried viability of probiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that uses environmental stress to stimulate extracellular polymeric substance synthesis for self-encapsulation formation combined with the addition of cryoprotectants to enhance the freeze-dried survival of probiotics. This could be a novel approach in improving the viability of probiotic strains for various applications.

Correlation Between the Amount of Extracellular Polymeric Substances and the Survival Rate to Freeze-Drying of Probiotics.

To demonstrate that the amount of extracellular polymeric substances (EPS) and the freeze-dried viability of probiotics are correlated. Three strains of probiotics including Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Bifidobacterium bifidum were subjected to environmental challenges, such as temperature, pH, and carbon dioxide. The results indicated that the challenges could stimulate the EPS synthesis of the probiotics. The experimental correlation between the amount of synthesized EPS and the freeze-dried survival rate was also analyzed, and the viability of each of the three strains was represented by the following functions in which the equation of L. plantarum is y = - 0.0336x2 + 2.7059x - 14.849 with R2 = 0.9699, the B. bifidum's equation is y = - 0.0554x2 + 2.6243x - 13.654 with R2 = 0.9554, and the L. acidophilus's one was y = 0.0346x2 + 0.5862x - 9.1339 with R2 = 0.9733. This could be a new approach to determining the freeze-dried viability of probiotic strains based on the measured EPS content.

Expression of genes involved in exopolysaccharide synthesis in Lactiplantibacillus plantarum VAL6 under environmental stresses.

Environmental factors can alter exopolysaccharide biosynthesis in lactic acid bacteria (LAB). To further clarify this potential relationship, the mRNA expression of genes involved in exopolysaccharide synthesis such as glmU, pgmB1, cps4E, cps4F, cps4J, and cps4H in Lactiplantibacillus plantarum VAL6 under different conditions including temperature, pH, sodium chloride (NaCl), and carbon dioxide (CO2) intensification culture was studied. The transcriptomic data revealed that the exposure of L. plantarum VAL6 at pH 3 increased the expression level of cps4H but decreased the expression levels of pgmB1 and cps4E. Under pH 8, cps4F and cps4E were significantly upregulated, whereas pgmB1 was downregulated. Similarly, the expression levels of cps4F, cps4E, and cps4J increased sharply under stresses at 42 or 47 °C. In the case of NaCl stress, glmU, pgmB1, cps4J, and cps4H were downregulated in exposure to NaCl at 7 and 10% concentrations while cps4E and cps4F were upregulated at 1 h of 10%-NaCl treatment and at 5 h of 4%-NaCl treatment. Remarkably, CO2 intensification culture stimulated the expression of all tested genes. In addition, simultaneous changes in expression of cps4E and cps4F under environmental challenges may elicit the possibility of an association between the two genes. These findings indicated that the expression level of eps genes is responsible for changes in the yield and monosaccharide composition of exopolysaccharides under environmental stresses.

Characterisation of Phosphate Accumulating Organisms and Techniques for Polyphosphate Detection: A Review.

Phosphate minerals have long been used for the production of phosphorus-based chemicals used in many economic sectors. However, these resources are not renewable and the natural phosphate stocks are decreasing. In this context, the research of new phosphate sources has become necessary. Many types of wastes contain non-negligible phosphate concentrations, such as wastewater. In wastewater treatment plants, phosphorus is eliminated by physicochemical and/or biological techniques. In this latter case, a specific microbiota, phosphate accumulating organisms (PAOs), accumulates phosphate as polyphosphate. This molecule can be considered as an alternative phosphate source, and is directly extracted from wastewater generated by human activities. This review focuses on the techniques which can be applied to enrich and try to isolate these PAOs, and to detect the presence of polyphosphate in microbial cells.

Biochemical Engineering Approaches for Increasing Viability and Functionality of Probiotic Bacteria.

The literature presents a growing body of evidence demonstrating the positive effect of probiotics on health. Probiotic consumption levels are rising quickly in the world despite the fluctuation of their viability and functionality. Technological methods aiming at improving probiotic characteristics are thus highly wanted. However, microbial metabolic engineering toolbox is not available for this kind of application. On the other hand, basic microbiology teaches us that bacteria are able to exhibit adaptation to external stresses. It is known that adequately applied sub-lethal stress, i.e., controlled in amplitude and frequency at a given stage of the culture, is able to enhance microbial robustness. This property could be potentially used to improve the viability of probiotic bacteria, but some technical challenges still need to be overcome before any industrial implementation. This review paper investigates the different technical tools that can be used in order to define the proper condition for improving viability of probiotic bacteria and their implementation at the industrial scale. Based on the example of Bifidobacterium bifidum, potentialities for simultaneously improving viability, but also functionality of probiotics will be described.

Scalable temperature induced stress for the large-scale production of functionalized Bifidobacteria.

The application of sub-lethal stresses is known to be an efficient strategy to enhance survival of probiotic bacteria during drying processes. In this context, we previously showed that the application of heat stress upon the entry into stationary phase increased significantly the viability of Bifidobacterium bifidum. However, this heat shock has been considered only in small-scale bioreactor and no information is available for a possible scaling-up strategy. Five different operating scales (0.2, 2, 20, 200 and 2000 L) have thus been tested and the results showed that the viability of B. bifidum increases from 3.15 to 6.57 folds, depending on the scale considered. Our observations pointed out the fact that the heat stress procedure is scalable according to the main outcome, i.e., increases in cell viability, but other factors have to be taken into account. Among these factors, dissolved carbon dioxide seems to play a significant role, since it explains the differences observed between the test performed at laboratory scale and in industrial conditions.

Halophilic lactic acid bacteria - Play a vital role in the fermented food industry.

Halophilic lactic acid bacteria have been widely found in various high-salt fermented foods. The distribution of these species in salt-fermented foods contributes significantly to the development of the product's flavor. Besides, these bacteria also have the ability to biosynthesize bioactive components which potentially apply to different areas. In this review, insights into the metabolic properties, salt stress responses, and potential applications of these bacteria have been have been elucidated. The purpose of this review highlights the important role of halophilic lactic acid bacteria in improving the quality and safety of salt-fermented products and explores the potential application of these bacteria.

Advances in Microbial Exopolysaccharides: Present and Future Applications.

Microbial exopolysaccharides (EPSs) are receiving growing interest today, owing to their diversity in chemical structure and source, multiple functions, and immense potential applications in many food and non-food industries. Their health-promoting benefits for humans deserve particular attention because of their various biological activities and physiological functions. The aim of this paper is to provide a comprehensive review of microbial EPSs, covering (1) their chemical and biochemical diversity, including composition, biosynthesis, and bacterial sources belonging mainly to lactic acid bacteria (LAB) or probiotics; (2) their technological and analytical aspects, especially their production mode and characterization; (3) their biological and physiological aspects based on their activities and functions; and (4) their current and future uses in medical and pharmaceutical fields, particularly for their prebiotic, anticancer, and immunobiotic properties, as well as their applications in other industrial and agricultural sectors.

Synbiotic Agents and Their Active Components for Sustainable Aquaculture: Concepts, Action Mechanisms, and Applications.

Aquaculture is a fast-emerging food-producing sector in which fishery production plays an imperative socio-economic role, providing ample resources and tremendous potential worldwide. However, aquatic animals are exposed to the deterioration of the ecological environment and infection outbreaks, which represent significant issues nowadays. One of the reasons for these threats is the excessive use of antibiotics and synthetic drugs that have harmful impacts on the aquatic atmosphere. It is not surprising that functional and nature-based feed ingredients such as probiotics, prebiotics, postbiotics, and synbiotics have been developed as natural alternatives to sustain a healthy microbial environment in aquaculture. These functional feed additives possess several beneficial characteristics, including gut microbiota modulation, immune response reinforcement, resistance to pathogenic organisms, improved growth performance, and enhanced feed utilization in aquatic animals. Nevertheless, their mechanisms in modulating the immune system and gut microbiota in aquatic animals are largely unclear. This review discusses basic and current research advancements to fill research gaps and promote effective and healthy aquaculture production.

Investigation of botulism in free-range duck farming in the Mekong Delta, Vietnam.

Background: One of the most common diseases in free-range ducks in the Mekong Delta is botulism. Botulism is a poultry disease caused by botulinum exotoxin of Clostridium botulinum. Aim: To evaluate the prevalence of botulism in free-range ducks in the Mekong Delta and the risk of infection by determining the presence of C. botulinum in the farming environment. Methods: Research was carried out on 200 duck flocks with 187,050 individuals raised freely in the fields in the provinces of the Mekong Delta, including An Giang, Can Tho, Hau Giang, and Kien Giang. The ducks were diagnosed with botulism based on clinical symptoms. To demonstrate the presence of botulinum neurotoxins and identify serotype, samples of serum and/or gut were analyzed by mouse bioassay. Samples of soil (n = 600), water (n = 600), crabs (n = 216), and snails (n = 400) were taken from the grazing regions for C. botulinum analysis by PCR assay. Results: There were 1.19% (2,235/187,050) free-range ducks in the Mekong Delta positive for botulism. Clinical symptoms of botulism including limberneck, drooping eyelids-enlarged pupils, and leg paralysis were prevalent across free-range ducks, with the frequency of 87.92% (1,965/2,235), 90.07% (2,013/2,235), and 79.78% (1,783/2,235), respectively. The lesions of pulmonary edema-hemorrhage, hemorrhagic liver, and gas-producing intestines were common, accounting for 86.19% (362/420), 95.48% (401/420), and 92.14% (387/420), respectively. Botulin toxin type C was found in a considerable number of serum samples, accounting for 40.48% (51/126). Meanwhile, the percentage of serum samples containing botulin toxin types E and D was 28.57% (36/126) and 25.40% (32/126), respectively. Clostridium botulinum was detected in the farming environment specifically 17.5% (105/600) in soil, 19.67% (118/600) in water, 8.33% (18/216) in crabs, and 3.00% (12/400) in snails. Conclusion: The free-range ducks in the Mekong Delta were at high risk of botulism because of the latent presence of C. botulinum in the farming environment.

Synbiotics: a New Route of Self-production and Applications to Human and Animal Health.

Synbiotics are preparations in which prebiotics are added to probiotics to achieve superior performance and benefits on the host. A new route of their formation is to induce the prebiotic biosynthesis within the probiotic for synbiotic self-production or autologous synbiotics. The aim of this review paper is first to overview the basic concept and (updated) definitions of synergistic synbiotics, and then to focus particularly on the prebiotic properties of probiotic wall components while describing the environmental factors/stresses that stimulate autologous synbiotics, that is, the biosynthesis of prebiotic-forming microcapsule by probiotic bacteria, and finally to present some of their applications to human and animal health.

Response of Lactobacillus plantarum VAL6 to challenges of pH and sodium chloride stresses.

To investigate the effect of environmental stresses on the exopolysaccharide biosynthesis, after 24 h of culture at 37 °C with pH 6.8 and without sodium chloride, Lactobacillus plantarum VAL6 was exposed to different stress conditions, including pH (pHs of 3 and 8) and high sodium chloride concentration treatments. The results found that Lactobacillus plantarum VAL6 exposed to stress at pH 3 for 3 h gives the highest exopolysaccharide yield (50.44 g/L) which is 6.4 fold higher than non-stress. Under pH and sodium chloride stresses, the mannose content in exopolysaccharides decreased while the glucose increased in comparison with non-stress condition. The galactose content was highest under stress condition of pH 8 meantime rhamnose content increased sharply when Lactobacillus plantarum VAL6 was stressed at pH 3. The arabinose content in exopolysaccharides was not detected under non-stress condition but it was recorded in great amounts after 3 h of stress at pH 3. In addition, stress of pH 8 triggered the mRNA expression of epsF gene resulting in galactose-rich EPS synthesis. According to our results, the stresses of pH and sodium chloride enhance the production and change the mRNA expression of epsF gene, leading to differences in the monosaccharide composition of exopolysaccharides.

Exopolysaccharide production by lactic acid bacteria: the manipulation of environmental stresses for industrial applications.

Exopolysaccharides (EPSs) are biological polymers secreted by microorganisms including Lactic acid bacteria (LAB) to cope with harsh environmental conditions. EPSs are one of the main components involved in the formation of extracellular biofilm matrix to protect microorganisms from adverse factors such as temperature, pH, antibiotics, host immune defenses, etc.. In this review, we discuss EPS biosynthesis; the role of EPSs in LAB stress tolerance; the impact of environmental stresses on EPS production and on the expression of genes involved in EPS synthesis. The evaluation results indicated that environmental stresses can alter EPS biosynthesis in LAB. For further studies, environmental stresses may be used to generate a new EPS type with high biological activity for industrial applications.