Probiotic Cultures Research Articles

Bioprocessing strategies for enhanced probiotic extracellular vesicle production: culture condition modulation.

Probiotic extracellular vesicles are biochemically active structures responsible for biological effects elicited by probiotic bacteria. Lactobacillus spp., which are abundant in the human body (e.g., gut), are known to have anti-inflammatory and antimicrobial properties, and are commonly used in food products, supplements, and in discovery research. There is increasing evidence that Lactobacillus-derived extracellular vesicles (LREVs) have potent immunomodulatory capacity that is superior to probiotics themselves. However, key mechanistic insights into the process that controls production and thus, the function of LREVs, are lacking. Currently, it is unknown how the probiotic culture microenvironment orchestrates the type, yield and function of LREVs. Here, we investigated how multifactor modulation of the biomanufacturing process controls the yield and biological functionality of the LREVs. To achieve this, we selected Lacticaseibacillus rhamnosus as the candidate probiotic, initially cultivated under traditional culture conditions, i.e., 100% broth concentration and pH 5.5. Subsequently, we systematically modified the culture conditions of the probiotic by adjusting three critical process parameters: (1) culture medium pH (pH 3.5, 5.5 and 7.5), (2) growth time (48 and 72h), and (3) broth concentration (50% and 10% of original broth concentration). EVs were then isolated separately from each condition. The critical quality attributes (CQA) of LREVs, including physical characteristics (size, distribution, concentration) and biological composition (protein, carbohydrate, lipid), were analysed. Functional impacts of LREVs on human epidermal keratinocytes and Staphylococcus aureus were also assessed as CQA. Our findings show that the production of LREVs is influenced by environmental stresses induced by the culture conditions. Factors like broth concentration, pH levels, and growth time significantly impact stress levels in L. rhamnosus, affecting both the production and composition of LREVs. Additionally, we have observed that LREVs are non-toxicity for keratinocytes, the major cell type of the epidermis, and possess antimicrobial properties against S. aureus, a common human skin pathogen. These properties are prerequisites for the potential application of EVs to treat skin conditions, including infected wounds. However, the functionality of LREVs depends on the culture conditions and stress levels experienced by L. rhamnosus during production. Understanding this relationship between the culture microenvironment, probiotic stress response, and LREV characteristics, can lead to the biomanufacturing of customised probiotic-derived EVs for various medical and industrial applications.

The effect of a synbiotic feed additive on the body of young pigs

The publication presents the results of the effect on the body of young pigs reared with a synbiotic feed additive. The synbiotic product consists of two components: lyophilized probiotic cultures isolated from the digestive system of pigs, and a vegetable prebiotic filler. These ingredients are combined into a dry mixture to form a synbiotic with pro- and prebiotic properties. The effect of the additive on the viability of livestock, live weight and its growth, indicators of feed consumption were studied, and the results of general and biochemical blood analysis of experimental animals were analyzed. The study showed that the inclusion of synbiotics in the diet of young pigs does not cause negative effects and is well tolerated by animals. In pigs receiving the supplement, an improvement in a number of indicators was noted: an increase in body weight gain, an increase in survival rate, as well as a decrease in feed consumption per unit of production. A blood test revealed a positive effect on hematopoiesis, in particular, on the formation of red blood cells and general hematopoiesis. In addition, an intensification of protein metabolism was observed. These results indicate a beneficial effect of synbiotic on the general condition and productivity of young pigs.

Исследование возможности обогащения шоколадных и снековых изделий пробиотическими культурами

Nutritional prevention of metabolic disorders, by introducing functional and specialized foods into the daily diet of the population, is one of the effective ways to solve the problem of reducing the growth of chronic non-communicable diseases. In this regard, today the awareness of the population in the field of the relationship between nutrition and health is growing, reflected in the eating behavior of the inhabitants. The article presents information on the enrichment of chocolate products with probiotic microorganisms. The indicators of viability and enzyme activity of various associations of probiotic cultures, including strains of Streptococcus lactis SL215, Streptococcus salivarius sp. thermophilus ST14, Lactobacillus acidophilus LA72, Lactobacillus delbrueckii sp bulgaricus LB50, Lactobacillus casei LC005, Lactobacillus rhamnosus LR112, Bifidobacterium bifidum BB79 in various combinations, were analyzed. As a result of the fortification of chocolate products with lyophilized powder of probiotic microorganisms with the species composition of Lactobacillus casei LC005, Lactobacillus rhamnosus LR112 (at a ratio of 1: 1), it was found that the introduction of lactic acid bacterias did not have significant differences in the organoleptic and rheological parameters of the product. The evaluation of the physico-chemical parameters of finished products indicates a high nutritional and biological value of chocolate.

Development and characterization of a novel flavored functional fermented whey-based sports beverage fortified with Spirulina platensis

Spirulina platensis, a microalga known for its exceptional nutritional value, especially its bioactive compounds and protein content, holds promise for incorporation into functional food products. Ricotta cheese whey is a byproduct of the production of ricotta cheese that is difficult to use in industries due to its low pH and less favorable processing qualities. This research aimed to create a unique fermented ricotta cheese whey-based beverage supplemented with various Spirulina powder concentrations (0.25 %, 0.5 %, and 0.75 % w/w) cooperated with a mixture of lemon and peppermint juice 10 % and fermented by probiotic (ABT) culture. The physicochemical, rheological, bioactive compounds, microbiological, and sensory properties were evaluated over a storage period of 21 days at cold storage. Spirulina-fermented whey-based beverages with a mixture of lemon and peppermint juice increased the concentration of vitamins, minerals, antioxidants, and total phenolic compounds in the final product. The count of probiotic bacteria in all fermented beverage samples exceeded 7 log CFU/mL throughout storage, indicating that the fermented beverage kept its probiotic properties. The addition of 0.5 % Spirulina significantly improved the final product's structural qualities and sensory acceptance.

Eşek sütünden yapılan fermente kımız

The study aimed to expand the range of dairy products by developing a recipe and technology for a fermented milk drink made from donkey milk with unique properties. Additional functionality is provided by the introduced probiotic cultures and phytogenic raw materials. Two functional drinks were proposed as experimental products: Koumiss-1 - koumiss with added probiotics and peony tincture, and Koumiss-2 - probiotic koumiss with added dried burdock roots. The initial raw material, donkey milk, was collected in the farm of the Ala Too State Farm located at an altitude of 800 m above sea level, in the Chui Valley of the Kyrgyz Republic. The standard methods of physicochemical, microbiological and organoleptic analyses were used for the study. The fermentation intensity (Is) and fermentation coefficient (δ) were determined for each case based on the data on the change in active and titratable acidity during the fermentation of milk with koumiss ferment. In the sample of koumiss-1 with the addition of peony tincture δ appeared in the first 3 hours, and δ in koumiss-2 was closer to 6 hours. Based on organoleptic evaluations, the developed drinks exhibit a pleasant sour-milk aroma and taste. The consistency is liquid with a slightly foamy texture, while the drink with added burdock roots has a pleasant creamy flavor. Lactic acid bacteria and yeast are similar to the specifications found in natural koumiss made from mare's milk. Throughout the experiment, sour-milk beverages were successfully produced, and they can potentially to fill the market niche for functional products with medicinal properties.

THE INFLUENCE OF NUTRIENTS AND LACTULOSE PREBIOTIC ON THE FERMENTATION PROCESSES IN THE TECHNOLOGY OF RECEIVING A FUNCTIONAL FOOD PRODUCT WITH AN INCREASED TITER OF VIABLE LACTIC ACID BACTERIA

A study of the influence of biologically active substances (fat-soluble vitamins, omega-3 fatty acids and vitamin D, as well as water-soluble vitamin C), prebiotic lactulose on the titer of viable cells of lactic acid bacteria was carried out for the further use of the obtained research results in the biotechnology of functional food products, obtained thanks to the successful course fermentation processes. Experiments have proven the expediency of using biologically active substances as stimulators of the growth of lactic acid bacteria, which contribute to the increase of biomass and support of their vital activity for 192 hours. Research has established that at the end of the experiment (192 hours), the highest titer of lactic acid bacteria was recorded in samples 1, 2, 3, 4, to which the prebiotic lactulose was added in the amount of 1, 3, 5 and 7 %, respectively. This is due to the fact that the prebiotic acts as a specific substrate that is used by the symbiosis of pure cultures of lactic acid bacteria for a long time, unlike milk sugar. Analyzing the obtained data, it is indicated that thanks to the addition of fat-soluble nutrients omega-3 fatty acids and vitamin D, as well as water-soluble vitamin C, there is a significant increase in the titer of viable cells of probiotic cultures at 6 and 24 hours of the experiment, although at the end of the experiment (by 192 hour) in these samples, an insignificant decrease of this indicator was recorded — from 10 to 8.9 log cells/ml. In the control sample (without the addition of biologically active substances and lactulose prebiotic), a significant decrease in the titer of viable cells of lactic acid bacteria was noted, compared to the test samples to which nutrients were added. The titer of lactic acid bacteria cells in the control sample was reduced by 192 hours from 8.5 to 5.5 log cells/ml. It is recommended to use the studied nutrients in the indicated concentrations to stimulate the growth and maintain the viability of lactic acid bacteria cultures in the technology of production of functional food products.

Biologically Active Complex Based on Fermented Fiber to Normalize and Ensure the Vital Microflora Activity

Nowadays, the intestinal microflora plays the important role in ensuring normal metabolism, no doubt. Therefore, biotechnological products and its metabolites are of particular importance. A man developed biotechnological product in the form of a biologically active additive based on fermented fiber (a mixture of dietary fibers bacterized with probiotic cultures of Bifidobacterium animalis, Bifidobacterium adolescentis, Lactobacillus acidophilus). The probiotic feature is the ability to ferment dietary fibers while staying alive in a specialized product. The formulation includes dietary fibers that stimulate the growth and bacteria reproduction, specifically, forming a target intestinal biotope. Along with traditional quality and safety research methods, the researchers used the method of high-performance liquid chromatography to determine the short-chain fatty acids content, playing a key role in the metabolic body processes, in fermented fiber. The formulation of the developed additive contains ingredients that form its properties. The determined regulated quality indicators are the following (in 30 g): dietary fiber – at least 20,000 mg; polypructosans – at least 2,200 mg; thiamine (B1) – 1.4 mg; riboflavin (B2) – 1.6 mg; pantothenic acid (B5) – 6 mg; pyridoxine (B6) – 2 mg; chromium – 50 mcg. The authors run the safety dietary supplements assessment, enabling to determine the shelf life – 12 months at a temperature of 25 °C and a relative humidity of no more than 60 %. Evidence-based medical studies confirm the functional orientation and effectiveness of the developed product. A man took the developed supplement one serving (15 g) twice a day (in the morning 30 minutes before meals and in the evening before bedtime). The administration course was 30 days. The data obtained indicate a positive impact of the biocomplex on the microbiota restoration, the local intestinal syndrome effect and the general condition of the body.

The Use of probiotics in fermenting food wastes for production of black soldier fly larvae (Hermetia illucens L.; Diptera: Stratiomyidae)

Larvae of the black soldier fly (BSF) can process a wide variety of organic wastes and convert them into larval body primarily consisting of protein. Production of larvae with high-quality protein content is affected the rearing system, one of them is quality of feeding substrates. This study was aimed to evaluate the effect of food wastes fermented by probiotic culture as the feeding substrates for producing the BSF larvae. The probiotic cultures used were Lactiplantibacillus plantarum E2 and Limosilactobacillus fermentum F5, while the food wastes were derived from a seafood restautrant. Four rearing substrates were compared: food wastes fermented by Lactiplantibacillus plantarum E2 (P1), food wastes fermented by Limosilactobacillus fermentum F5 (P2), food wastes fermented by consortium (Lactiplantibacillus plantarum E2 + Limosilactobacillus fermentum F5) (P3), and food wastes without probiotics (P0). Before the feeding substrates were applied, the food wastes were fermented by the probiotics for 10 days. Six-day-old BSF larvae were reared for 14 days on each fermented substrate. The larval growth performance observed were fresh weight, length and width, while the nutrient content of the larvae was protein and fat content. The waste reduction index was also measured. The data showed that the best larval growth performance was shown in P3 substrate, with fresh weight of 287.27 g, larva length and width of 19.93 mm and 3.53 mm, respectively. In P3 substrate also could produce the larvae with the best nutrient content, with protein and fat content by 12.53% and 16.09%, respectively. The waste reduction index value of P3 substrate was 6.12%. The duration of BSF larvae rearing required to be extended to obtain an increased average protein content. This finding of this study could be helpful for improving the quality of BSF larvae by applying consortium of probiotic cultures in fermenting the feeding substrates.

Investigation of aroma profiles, textural, rheological, and sensorial qualities of yogurts with various starter cultures and goat–cow milk combinations

Abstract In the present study, aroma components, textural, rheological and sensory qualities of yogurts produced using YC-350 and ABT-2 starter cultures and combinations of goat/cow milk were investigated during the storage period. For this purpose, aroma components were analyzed by GC-MS, textural analyses by TA.XTplus Texture Analyzer and rheological analyses by Anton Paar MCR 102 rheometer. The effects of goat/cow milk combinations and variety of starter culture on aroma components, textural, rheological and sensory properties of yogurt samples were found significant (p < 0.01). Also, the storage period significantly (p < 0.01) affected all parameters tested except cohesiveness, resilience, and appearance. While it was found that the use of cow milk in the experimental yogurt samples increased the acetaldehyde content, it was found that it had varying effects on the diacetyl content, acetoin content, and textural and rheological analysis parameters. It was determined that the content of diacetyl and acetoin reduced with the progression of the storage times. In the sensory evaluations, the use of goat milk had positive effects on the parameters of appearance and consistency by spoon, and the use of probiotic culture (ABT-2) had positive effects on the parameters of consistency by mouth and overall acceptability.

Synbiotic supplements as antibiotic alternatives in broiler diets

Antibiotics can contribute to bacterial resistance, posing a significant public health concern. Synbiotics represent an effective alternative to antibiotics, promoting the balance of intestinal microbiota and creating a conducive environment for beneficial bacteria growth. This study aimed to assess the utility of the synbiotic supplement (+Poultry) as a substitute for antibiotics in broiler diets from 1 to 42 days of age. We evaluated its impact on performance during two stages: 1 to 21 days and 1 to 42 days, digestibility coefficients, metabolizable energy, duodenum and jejunum morphometry, carcass yield, and hematological parameters. A total of 440 Ross 308 strain broiler chicks, both males and females, were accommodated in 2m² cages from day 1 to day 42. The birds were randomly assigned to four treatments, with each treatment having five replications consisting of 22 birds, thus forming 20 experimental plots. The treatment groups were as follows: Basal diet (BD) without the addition of antibiotics or synbiotics, BD with 150ppm zinc bacitracin (BCZ), BD with 0.5kg/t of synbiotic, and BD with 1kg/t of synbiotic. The synbiotic supplement (+Poultry) used in this experiment, provided by Nutri+, contained amino acids, minerals, probiotic culture, prebiotic additives, and flavoring additives. Throughout the experiment, all birds were subjected to a weekly sanitary challenge, which involved using reused litter and water contaminated with litter. Treatment effects were observed on feed conversion (P = 0.0100) of broilers aged 1 to 21 days, indicating improved feed conversion in broilers consuming BD + 1kg/t synbiotic compared to BD alone. Consequently, we recommend the incorporation of a synbiotic supplement at a rate of 1kg/t as a viable antibiotic replacement for chickens aged 1 to 21 days. The tested synbiotic supplement shows promise as an antibiotic alternative in broiler diets at all life stages, maintaining both performance and bird health.

Биотехнология производства мягких сыров из смеси коровьего и козьего молока с функциональными ингредиентами

The aim of this research is to develop a biotechnology of soft cheeses with func-tional ingredients based on a mixture of cow’s and goat’s milk taken in a ratio of 60:40, subjected to acoustic cavitation at 75 kHz and an exposure time of 17 min. To achieve the research aim, the tasks of selecting and determining the number of functional ingredients, determining the technological parameters of the production of soft cheeses with their use were solved. As a result, the biotechnology of two types of soft cheeses «Dieticheskij» and «Funkcionalnyj» has been developed. In the biotechnology of soft cheese «Dieticheskij», a complex preparation «Bivestin-lacto» containing cultures of probiotics Bifidobacterium longum MS-42, Lactoba-cillus plantarum 8PA-3 and dietary fiber «Citri-Fi» of medium grinding was used as a functional ingredient. Soft cheese «Funkcionalnyj» is enriched with a biocomponent containing probiotic cultures of Lactobacillus acidophilus La-5, Lactobacillus delbrueckii subsp. Bulgaricus, Bifidobacterium lactis, Streptococcus thermophilus, as well as «Selexen» – a source of selenium, «Curcumin» in micellar form and dietary fiber «Citri-Fi». Their organoleptic, chemical, microbiological parameters, as well as nutritional, energy value and shelf life have been determined.

Normalization of Deviant Behavior in Muc2+/+ Mice through Dietary Incorporation of Bacillus subtilis Spores

It is well-known that the intestinal microbiota influences the host's health, including the activities of the central nervous system. This phenomenon is known as the brain-gut axis. Mental and emotional disorders are now fairly widespread. Many of them are lifelong, and medications frequently cause side effects. Not surprisingly, researchers have been drawn to the study of probiotic bacterial cultures as an adjunct or primary therapy for psycho-emotional illnesses. There is a lot of evidence that bacterial strains from the genera Bifidobacterium and Lactobacillus have a favorable influence on the activities of the central nervous system. However, the effect of Bacillus spp. on the behavior is not clearly known. In contrast to Muc2-/-, the wild type mice (Muc2+/+) have normal gut barrier function, but the intestinal microbiota and behavior are similar to their mutant siblings Muc2-/- mice. Similar to the mutant sibling, Muc2+/+mice exhibit greater locomotor activity in the open field and light-dark tests, as well as decreased anxiety in the open field test, light-dark test, and marble burying tests. The addition of kanamycin-resistant strain Bacillus subtilis BS20 spores to the diet of Muc2+/+ mice for 3 weeks at 107 CFU/g led to a change in behavior and cytokine levels in the gut compared to control C57BL/6 mice. Furthermore, a drop in serotonin and a rise in tyrosine were identified in the blood serum. Changes in cytokines, serotonin, and tyrosine levels may mediate the normalizing impact of B. subtilis spores on Muc2+/+ mice behavior.

METABOLIC CHANGES IN BURNS DURING ORAL ADMINISTRATION OF AEROCOCCUS VIRIDANS IN UNCOMFORTABLE TEMPERATURE CONDITIONS

The organism of farm animals is in an inextricable connection with the environment, with which there is a constant exchange of substances, energy and information, that is, it is an open metabolic system that depends on the conditions of existence. Therefore, it is very important to create the most favorable technologies for exploiting the physiological capabilities of animals in conditions of physiological well-being and epidemiological well-being. This is the foundation for obtaining high-quality and bio-safe animal husbandry products. One of the common negative factors of violations of the physiological needs of the animal body is the effect of low-intensity permanent stress of multivalent etiology. During the development of negative pathophysiological disorders, the quantitative and qualitative composition of the microbiota of the large intestine changes, as a dynamic microbial community of transient and resident populations that develop according to the chemo-static principle of functioning. Restoration and correction of the microbiota of the large intestine and transformation of the disturbed equilibrium of the interpopulation ratio is an effective means of eliminating the negative effects of stress on the animal body. The resident probiotic microflora is represented by a wide variety of prokaryotic microbiota and many reference strains are used for therapeutic and prophylactic purposes. Aerococcus viridans is very promising and effective, which lives in various biotopes of the macro-organism and is an indicator of the physiological well-being of the organism, because it is sensitive to toxic compounds and antimicrobial substances, but with permanent use it exhibits pronounced antagonistic properties against transient microflora with potentially pathogenic properties due to inhibition of their colonization abilities and exhibits detoxification potentials. Ants were used to simulate low-intensity permanent stress. as heat-loving animals sensitive to uncomfortable temperatures of the external environment and placed them in a household refrigerator for ten days at a temperature of 6-8 oC. Probiotic culture of production strain BI-07 Aerococcus viridans in a dose of 2×109 CFU/cm3 was administered orally as an anti-stress factor. A similar experiment was conducted in comfortable conditions, that is, at a temperature of 20-22 oC. Metabolic changes in the body of ants were recorded during a laboratory blood test. Probiotic culture does not affect the normergic processes of the macro-organism and does not change indicators beyond the physiological norm, but when used in conditions of low-intensity temperature stress, it has a positive effect on the adaptation-compensatory mechanisms of the macro-organism and shows protective potential in relation to non-resident microbiota.

Improvement of the probiotic cultures method isolation from freshly extracted honey

The study of probiotic microorganisms isolated from bee honey has recently become widespread in the world and is of great interest both for the prevention and treatment of bee diseases, and as a source of probiotics for the design of drugs useful for humans and animals. However, no studies have been conducted to study the duration of probiotic bacteria storage in honey after it has been pumped out of the combs. Three types of honey (acacia, sunflower and multi-herb) were studied to determine the intensity of growth of probiotic bacteria on the first, second, third and fourth day after pumping, as well as after storing honey for 6 months in sealed honeycombs. Honey samples were prepared in dilution with MRS broth, sown on a solid nutrient medium, the results were recorded after 48 hours by counting colonies of different species. Selected cultures were characterized by cultural, morphological and biochemical properties. It was established that bacteria are released during the first 3 days after honey is pumped out, on the other hand, on the 4th day of storage, there is no growth of probiotic bacteria, which indicates the impossibility of obtaining them. The prospect of using honey of any different botanical origin for the isolation of probiotic bacteria has been proven. A study of honey stored in sealed honeycombs for 6 months confirmed the hypothesis that normal flora bacteria with probiotic properties are stored in honeycombs for a long time and can be used for their isolation, identification and further use. In the course of the study, the method of isolating bacteria of the normal flora of the intestine of bees, namely Lactobacillus plantarum, Bifidobacterium bifidum and Enterococcus faecium from freshly extracted honey, was improved and developed. Key words: normal flora, probiotic bacteria, freshly squeezed honey, Lactobacillus plantarum, Bifidobacterium bifidum, Enterococcus faecium.

Effect of two lyoprotectants on the survival rate and storage stability of freeze-dried probiotic lactic acid bacterial strains

Lactic acid bacteria are very important for biotechnology, pharmaceutical and food industry, and are extensively used as probiotic products and starter cultures. Freeze-drying is a widely used and effective process for preservation of lactic acid bacteria in a dried form. To ensure stability and maintain biological activity of lactic acid bacteria, suitable protective media formulations should be used during the freeze-drying process. Twelve oral strains with functional and probiotic properties were subjected to freeze-drying and assessed for viability after the process and after long-term storage. Two lyoprotective media with different composition were used and the freeze-drying process was performed, where retained stability of the studied strains in both lyoprotectors was reported. Viability after four and eight months of storage was assessed, showing that the strains maintained high viability. The two lyoprotective media ensured the obtaining of freeze-dried samples with preserved viability for all studied strains of lactic acid bacteria. Four of the tested strains, L. fermentum N 2 and TC 3-11, L. delbrueckii subsp. lactis VG 2, and W. confusa NN 1 showed well preserved stability and viability after freeze-drying and long-term storage and, prepared in freezedried formulations, are suitable to be included in the omposition of probiotic products for oral healthcare.

The role of a universal microbiological ferment with probiotic action in changing the blood picture of calves in preweaning period

The use of probiotic feed additives is known in animal science and practice; they are an alternative way to replace antibiotics in animal husbandry. The use of probiotic drugs signifi cantly increases the natural resistance of the animal body, maintains a high level of immune status and reduces the risk of infectious diseases in cattle and farm animals in general. The purpose of the research was to study the eff ect of feeding a universal microbiological ferment with probiotic action Biolax-U in the diets of calves during the preweaning period in combination with a hygienic bedding desiccant Drytall Fresh. In order to conduct the research 2 groups (control and experimental) of clinically healthy calves of Black-and-White breed in the preweaning period per 15 heads in each were formed by the method of analogues, taking into account age, live weight and breed. Calves of the compared groups received milk and basic feed according to the calf feeding scheme used on the farm. At the same time, the calves of the experimental group additionally received the universal microbiological ferment with probiotic action Biolax-U per 10 ml each morning and evening feeding, i.e. in two steps. The use of Biolax-U ferment had the positive eff ect on improving the morphological composition and biochemical parameters of blood. A comparative blood test after 90 days of feeding calves in the experimental group with the probiotic ferment culture Biolax-U revealed the increase in the concentration of erythrocytes by 12,20 %, hemoglobin by 10,58 % compared to the beginning of the experiment, and by 7,26 and 11,02 %, respectively, compared to animals in the control group. It has been established that the use of the Biolax-U ferment in the diet of calves during the preweaning period in combination with the hygienic bedding desiccant Drytall Fresh enhanced redox processes in the body of calves, stimulated erythropoiesis, improved oxygen supply to tissues and organs, increased the intensity of protein metabolism and protective functions body.

Gas Chromatography-Mass Spectrometry Profiling of Volatile Metabolites Produced by Some Bacillus spp. and Evaluation of Their Antibacterial and Antibiotic Activities.

Bacillus species produce different classes of antimicrobial and antioxidant substances: peptides or proteins with different structural compositions and molecular masses and a broad range of volatile organic compounds (VOCs), some of which may serve as biomarkers for microorganism identification. The aim of this study is the identification of biologically active compounds synthesized by five Bacillus species using gas chromatography coupled to mass spectrometry (GC-MS). The current study profoundly enhances the knowledge of antibacterial and antioxidant metabolites ensuring the unambiguous identification of VOCs produced by some Bacillus species, which were isolated from vegetable samples of potato, carrot, and tomato. Phylogenetic and biochemical studies were used to identify the bacterial isolates after culturing. Phylogenetic analysis proved that five bacterial isolates BSS12, BSS13, BSS16, BSS21, and BSS25 showed 99% nucleotide sequence similarities with Bacillus safensis AS-08, Bacillus cereus WAB2133, Bacillus acidiproducens NiuFun, Bacillus toyonesis FORT 102, and Bacillus thuringiensis F3, respectively. The crude extract was prepared from bacterial isolates to assess the antibiotic resistance potency and the antimicrobial potential against various targeted multidrug-resistant strains, including yeast strains such as Candida albicans, Candida krusei, and bacterial strains of Enterococcus hirae, Escherichia coli, Klebsiella aerogenes, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus group B, Streptococcus mutans, Shigella sonnei, Salmonella enteritidis, Serratia marcescens, Pseudomonas aeruginosa, and Proteus vulgaris. GC-MS analysis of bacterial strains found that VOCs from Bacillus species come in a variety of chemical forms, such as ketones, alcohols, terpenoids, alkenes, etc. Overall, 69 volatile organic compounds were identified from five Bacillus species, and all five were found to share different chemical classes of volatile organic components, which have a variety of pharmacological applications. However, eight antibacterial compounds with different concentrations were commonly found in all five species: acetoin, acetic acid, butanoic acid, 2-methyl-, oxime-, methoxy-phenyl, phenol, 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester, nonanoic acid, and hexadecanoic acid, methyl. The present study has demonstrated that bacterial isolates BSS25, BSS21, and BSS16 display potent inhibitory effects against Candida albicans, while BSS25, BSS21, and BSS13 exhibit the ability to restrain the growth and activity of Candida krusei. Notably, BSS25 and BSS21 are the only isolates that demonstrate substantial inhibitory activity against Klebsiella aerogenes. This disparity in inhibitory effects could be attributed to the higher concentrations of acetoin in BSS25 and BSS21, whereas BSS16 and BSS13 have relatively elevated levels of butanoic acid, 2-methyl-. Certainly, the presence of acetoin and butanoic acid, 2-methyl-, contributes to the enhanced antibacterial potential of these bacterial strains, in conjunction with other organic volatile compounds and peptides, among other factors. The biology and physiology of Bacillus can be better understood using these results, which can also be used to create novel biotechnological procedures and applications. Moreover, because of its exceptional ability to synthesize and produce a variety of different antibacterial compounds, Bacillus species can serve as natural and universal carriers for antibiotic compounds in the form of probiotic cultures and strains to fight different pathogens, including mycobacteria.

Alternative Fermented Soy-Based Beverage: Impact of Inulin on the Growth of Probiotic Strains and Starter Culture

The number of people with dietary restrictions on dairy products has increased significantly due to lactose intolerance/allergy or adoption of vegan diets. Organic acid-producing probiotics have been used in fermented beverages, such as those based on soy, with good results. Such molecules have in fact been described for their role in sensory analyses and benefits to human health. Therefore, this study suggested the evaluation of an alternative soy extract-based beverage that could act as a functional food. For this purpose, products and biomass concentrations were monitored throughout soy extract fermentation through acidification kinetics and cell count. The effect of inulin on the growth of the probiotic strains Bifidobacterium longum and Lactobacillus acidophilus in co-culture with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was evaluated (technical replicates). It was observed that the addition of inulin reduced the time of fermentation by L. acidophilus, while no statistically significant effect was observed in the post-acidification period. In B. longum fermentation, the process did not change in the presence of inulin, but there was a significant increase in viability and survival in the post-acidification period. Therefore, it can be concluded that the strains studied can be used in the formulation of soy-based drinks and that inulin positively influenced the viability of both probiotics in fermented drinks tested.

Functional foods are essential components of nutrition

The most common category of functional nutrition is fermented milk products, which normalize the intestinal microflora and increase the immune status of the human body. Fermented milk products are characterized by high biological and nutritional values. This means a high degree of balance in the amino acid composition of milk proteins, compared to the so-called ideal food protein, the amino acid composition of which meets the needs of the human body. In addition, the proteins of fermented milk products are well digested by proteolytic enzymes of the gastrointestinal tract. The main casein proteins are capable of being broken down equally well by proteolytic enzymes in their native and denatured states. The macroorganism and intestinal microflora are a relatively stable and balanced ecological system, the balance of which, on the one hand, is determined by the physiological and immunological characteristics of the macroorganism, on the other, by the species and quantitative composition of microbial associations and the diversity of their biological activity. In a normal physiological state, the relationship between the macroorganism and the microflora is symbiotic in nature, and the latter has a significant impact on the general immunity and natural resistance of the host to infections, takes an active part in the processes of digestion and the synthesis of various biologically active substances. In turn, the macroorganism regulates the composition of the intestinal microflora due to factors such as the acidity of gastric juice and the content of bile salts. Fermented dairy products are the main suppliers of probiotic microorganisms that contribute to the maintenance and restoration of human microbial ecology. Probiotic cultures that provide beneficial effects on the consumer’s body and normalize the composition and functions of the microflora of the gastrointestinal tract include such types of lacto- and bifidobacteria as Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium spp. (B. adolescentis, B. animalis ssp. lactis, B. bifidum, B. longum, B. breve).

Yoghurt quality when using probiotic starter cultures and vegetable filler

In order to increase the biological value of yogurt, increase and improve its assortment, it is necessary to continue the search and use of new additives and fillers of natural origin. Such fillers should be well combined with the milk base, contain mineral elements, vitamins and other biologically active substances. The article substantiates the use of bacterial starters and vegetable filler in yogurt technology. The results of studies on the use of dry bacterial starter cultures of direct application of the Ukrainian manufacturer of the “YOGURTON” trademark and the vegetable filler “Black currants dry extract of 30 % anthocyanins” in the production of yoghurt are presented. An evaluation of the prototypes showed that when using the Yoghurt Probio TM «YOGURTON» starter, the product had a more pronounced taste and smell. The time of clot formation, changes in titrated acidity, and organoleptic characteristics of yogurt with filler were studied. It was found that all samples of yogurt on organoleptic parameters meet the requirements of DSTU and high scores on the scoring system of fermented milk products. The tested starter cultures at a fermentation temperature range of 40 ± 2 °C form a clot for 6–8 hours with a titratable acidity of 75 °T. Yogurt with a content of dry currant powder in an amount of 1 % in terms of organoleptic characteristics turned out to be the best; with an increase in the content of currant powder in yogurt up to 1.5 %, an increase in the taste of currants and a decrease in the characteristic sour milk taste of the product were noted. High acidity gives the product a sour taste and smell, which reduces its organoleptic characteristics. The acidity of yogurt with a filler content of 1 % was 116 °T on the first day of storage and increased to 140 °T for ten days. During storage of yogurt at the temperature of +6 ± 2 °C there is a gradual increase in lactic acid microorganisms, which affects the organoleptic characteristics of yogurt. “Black currant extract dry 30 % anthocyanins” enriches yogurt with anthocyanins, flavonoids, organic acids, pectin, which improve the taste and quality of the product.