Characterization of Selected Microalgae Species as Potential Sources of Nutrients and Antioxidants.

Microalgae are exceptional organisms from a nutritional perspective, boasting an array of bioactive compounds that have long justified their incorporation into human diets. In this study, we explored the potential of five microalgae species: Nannochloropsis sp., Tetraselmis chuii, Chaetoceros muelleri, Thalassiosira weissflogii, and Tisochrysis lutea. We conducted comprehensive analyses of their nutritional profiles, encompassing protein content, individual amino acid composition, mineral and trace element levels, fatty acid profiles (including saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs)), polyphenol compositions, and vitamin B content. The antioxidant activity of the ethanolic extracts was evaluated using two methods: ABTS and DPPH radical scavenging assay. The total protein content of the microalgae ranged from 34.09 ± 0.39% to 42.45 ± 0.18%, with the highest concentration observed in T. weissflogii. Essential amino acids such as histidine, threonine, lysine, valine, isoleucine, leucine, phenylalanine, and methionine were present in concentrations ranging from 0.53 ± 0.02 to 12.55 ± 2.21 g/16 g N. Glutamic acid emerged as the most abundant amino acid, with concentrations ranging from 6.73 ± 0.82 to 12.55 ± 2.21 g/16 g N. Among the microalgae species, T. chuii exhibited the highest concentrations of calcium (Ca) and manganese (Mn), while C. muelleri showed prominence in magnesium (Mg), sodium (Na), and iron (Fe). T. weissflogii stood out for its potassium (K) content, and T. lutea contained notable amounts of copper (Cu), zinc (Zn), and lead (Pb). Regarding fatty acid profiles, Nannochloropsis sp. and T. chuii were predominantly composed of SFA, while C. muelleri and T. weissflogii were rich in MUFA. PUFAs dominated the fatty acid profile of T. lutea, which also exhibited the most diverse range of polyphenolic substances. We also analyzed the B vitamin content, with T. lutea displaying the highest concentrations of niacin (B3) and riboflavin (B2). Antioxidant activity was confirmed for all microalgae tested using DPPH and ABTS radical IC50 (mg/mL) converted to Trolox equivalent (TEAC). These findings underscore the substantial potential of the examined microalgae species as sources of biologically valuable substances characterized by rapid growth and relatively undemanding cultivation conditions.

Temperature, Salinity and Garlic Additive Shape the Microbial Community during Traditional Beetroot Fermentation Process.

Plant-based traditional fermented products are attracting a lot of interest in global markets. An example of them is beetroot leaven, which is valued for its high bioactive compound content. The variety of production recipes and the spontaneous nature of red beet fermentation favor its high diversity. This study aimed to analyze the impact of external factors-temperature, brine salinity, and garlic dose-on the beetroot fermentation and bacterial metapopulation responsible for this process. The research results confirmed the significant influence of the selected and analyzed factors in shaping the leaven physicochemical profile including organic acid profile and betalain content. Analysis of bacterial populations proved the crucial importance of the first 48 h of the fermentation process in establishing a stable metapopulation structure and confirmed that this is a targeted process driven by the effect of the analyzed factors. Lactobacillaceae, Enterobacteriaceae, and Leuconostocaceae were observed to be the core microbiome families of the fermented red beet. Regardless of the impact of the tested factors, the leaven maintained the status of a promising source of probiotic bacteria. The results of this research may be helpful in the development of the regional food sector and in improving the quality and safety of traditionally fermented products such as beetroot leaven.

Optimization and Modeling of Citrobacter freundii AD119 Growth and 1,3-Propanediol Production Using Two-Step Statistical Experimental Design and Artificial Neural Networks.

1,3-propanediol (1,3-PD) has a wide range of industrial applications. The most studied natural producers capable of fermenting glycerol to 1,3-PD belong to the genera Klebsiella, Citrobacter, and Clostridium. In this study, the optimization of medium composition for the biosynthesis of 1,3-PD by Citrobacter freundii AD119 was performed using the one-factor-at-a-time method (OFAT) and a two-step statistical experimental design. Eleven mineral components were tested for their impact on the process using the Plackett-Burman design. MgSO4 and CoCl2 were found to have the most pronounced effect. Consequently, a central composite design was used to optimize the concentration of these mineral components. Besides minerals, carbon and nitrogen sources were also optimized. Partial glycerol substitution with other carbon sources was found not to improve the bioconversion process. Moreover, although yeast extract was found to be the best nitrogen source, it was possible to replace it in part with (NH4)2SO4 without a negative impact on 1,3-PD production. As a part of the optimization procedure, an artificial neural network model of the growth of C. freundii and 1,3-PD production was developed as a predictive tool supporting the design and control of the bioprocess under study.

Hermetia illucens frass improves the physiological state of basil (Ocimum basilicum L.) and its nutritional value under drought.

To counterbalance the growing human population and its increasing demands from the ecosystem, and the impacts on it, new strategies are needed. Use of organic fertilizers boosted the agricultural production, but further increased the ecological burden posed by this indispensable activity. One possible solution to this conundrum is the development and application of more environmentally neutral biofertilizers. The aim of this study was to compare the effectiveness of two doses of Hermetia illucens frass (HI frass) with the commercial cattle manure in the cultivation of basil under drought. Soil without the addition of any organic fertilizer was used as a baseline control substrate for basil cultivation. Plants were grown with cattle manure (10 g/L of the pot volume) or HI frass at two doses (10 and 12.5 g/L). The health and physiological condition of plants were assessed based on the photosynthetic activity and the efficiency of photosystem II (chlorophyll fluorescence). Gas exchange between soil and the atmosphere were also assessed to verify the effect of fertilizer on soil condition. In addition, the mineral profile of basil and its antioxidant activity were assessed, along with the determination of the main polyphenolic compounds content. Biofertilizers improved the fresh mass yield and physiological condition of plants, both under optimal watering and drought, in comparison with the non-fertilized controls. Use of cattle manure in both water regimes resulted in a comparably lower yield and a stronger physiological response to drought. As a result, using HI frass is a superior strategy to boost output and reduce the effects of drought on basil production.

Influence of the Polymerization Modes on the Methacrylic Acid Release from Dental Light-Cured Materials-In Vitro Study.

The study focuses on the problem of lowering the pH around a composite filling concerning the polymerization modes and methacrylic acid release, which may affect not only the oral health but also the whole organism. A total of 90 specimens (30 of each: Filtek Bulk Fill, Evetric and Riva LC) were placed in 90 sterile hermetic polyethene containers with saline and incubated at 37 °C. Ten samples of each material were light-cured for 40 s with one of the three curing modes: full power mode (FPM), ramping mode (RM) and pulse mode (PM). The pH and methacrylic acid release evaluation were performed at the following time points: after 2 h and after 3, 7, 21 and 42 days from the specimen preparation. Regardless of light-curing mode, all used materials were characterized by a gradual elevation in methacrylic acid concentration. Only for Filtek Bulk Fill, increased methacrylic acid release was closely associated with lower pH. The choice of the polymerization mode has no significant influence on the methacrylic acid release. However, further research about composite light-curing is necessary to create the procedure algorithm, reducing the local and systemic complications associated with composite fillings.

Pasta with Kiwiberry (Actinidia arguta): Effect on Structure, Quality, Consumer Acceptance, and Changes in Bioactivity during Thermal Treatment.

In this study, kiwiberry lyophilizate (KBL) was incorporated into pasta at different levels (5%, 10%, and 15% w/w). Kiwiberry fruits' characteristics (ascorbic acid, carotenoids, phenolic compounds, and antioxidant activity determination) as well as physical (cooking properties, color, microscopic structure determination, texture, and water molecular dynamics analysis by low-field NMR) and chemical analyses (proximate composition phenolic compounds composition and antioxidant activity) of KBL-enriched pasta were investigated. The replacement of semolina with KBL in the production of pasta significantly changed its culinary properties. Results showed that the addition of KBL leads to a reduction in optimal cooking time and cooking weight (47.6% and 37.3%, respectively). Additionally, a significant effect of the KBL incorporation on the color of both fresh and cooked pasta was observed. A significant reduction in the L* value for fresh (27.8%) and cooked (20.2%) pasta was found. The KBL-enriched pasta had a different surface microstructure than the control pasta and reduced firmness (on average 44.7%). Low-field NMR results have confirmed that the ingredients in kiwiberry fruit can bind the water available in fresh pasta. The heat treatment resulted in increasing the availability of phenolic compounds and the antioxidant activity (64.7%) of cooked pasta. Sensory evaluation scores showed that the use of 5-10% of the KBL additive could be successfully accepted by consumers.

The Effect of Plant Additives on the Stability of Polyphenols in Dried Black Chokeberry (Aronia melanocarpa) Fruit.

Chokeberry fruit exhibits a high level of pro-health potential, associated with a significant amount of polyphenol content and antioxidant activity. The fruit is easily perishable and therefore needs to be processed in short order to ensure its availability throughout the year. For this purpose, the fruit is dried, which has an impact on the preservation of bioactive components such as polyphenols. In the study, chokeberry fruit was influenced by a suspension of spices, including clove and cinnamon, and subsequently dried using the freeze-drying, convection, and microwave radiation methods. Freeze-drying was concluded to be the most desirable method of drying, which preserves the largest amount of polyphenols while soaking fruit in a suspension of cinnamon or cloves accelerated the loss of anthocyanins during the storage period. The obtained test results may constitute a valuable source of information for manufacturers in designing new products with increased pro-health potential, whose properties result from the high content of polyphenols and other plant additives.

The Effect of Plant Additives on the Stability of Polyphenols in Cloudy and Clarified Juices from Black Chokeberry (Aronia melanocarpa).

Black chokeberry (Aronia melanocarpa) is a fruit with increasing popularity in consumption and processing. Recent research has strengthened the position of chokeberry as a source of phenolic compounds, antioxidants with high pro-health values, therefore it is important to investigate other substances protecting biologically active compounds during juice processing. This study was an attempt to reduce the loss of polyphenol in cloudy and clarified chokeberry juice by adding aqueous cinnamon and clove extracts. The results showed that the clarification of juices did not cause significant changes in the concentration of polyphenols. However, the addition of plant extracts prior to pasteurisation process influenced the content of phenolic compounds in the chokeberry juices. The main change in the composition of the chokeberry juices observed during storage was a result of the degradation process of anthocyanins. The research showed that, despite the common view about the beneficial effects of polyphenols and other compounds exhibiting mutual antioxidative potential, it is very difficult to inhibit the degradation process.

Interactions between fecal bacteria, bile acids and components of tomato pomace.

The tomato pomace obtained during processing as a residue of tomato processing from large industry. The interactions between tomato pomace and fecal bacteria, bile acids during in vitro digestion were studied. Digestion was carried out by using bioreactor in anaerobic conditions. Tomato pomace can significantly affect the count of fecal bacteria and the solubility of bile acids in in vitro digestion due to bonding ability of their proteins/peptides. The availability and use of bile acids does not only depend on the interactions between bile acids and bacteria, but also the interactions of bile acids with digested food components. Tomato pomace characterized high dietary fiber content and its fractions: 17.64-21.53% for cellulose and 13.48-18.63% for lignin. Given our results we supposed that fecal bacteria can use primary bile acids, as their source of energy in an environment where carbon availability is limited.

Group II intron-mediated deletion of lactate dehydrogenase gene in an isolated 1,3-propanediol producer Hafnia alvei AD27.

Our previous studies showed that glycerol fermentation by Hafnia alvei AD27 strain was accompanied by formation of high quantities of lactate. The ultimate aim of this work was the elimination of excessive lactate production in the 1,3-propanediol producer cultures. Group II intron-mediated deletion of ldh (lactate dehydrogenase) gene in an environmental isolate of H. alvei AD27 strain was conducted. The effect of the Δldh genotype in H. alvei AD27 strain varied depending on the culture medium applied. Under lower initial glycerol concentration (20 gL-1), lactate and 1,3-propanediol production was fully abolished, and the main carbon flux was directed to ethanol synthesis. On the other hand, at higher initial glycerol concentrations (40 gL-1), 1,3-propanediol and lactate production was recovered in the recombinant strain. The final titers of 1,3-propanediol and ethanol were similar for the recombinant and the WT strains, while the Δldh genotype displayed significantly decreased lactate titer. The by-products profile was altered upon ldh gene deletion, while glycerol utilization and biomass accumulation remained unaltered. As indicated by flow-cytometry analyses, the internal pH was not different for the WT and the recombinant Δldh strains over the culture duration, however, the WT strain was characterized by higher redox potential.

Effect of Wheat Dietary Fiber Particle Size during Digestion In Vitro on Bile Acid, Faecal Bacteria and Short-Chain Fatty Acid Content.

The influence of bile acid concentration on the growth of Bifidobacterium spp. and Lactobacillus spp. bacteria was demonstrated. Exposing these bacteria to the environment containing bile acid salts, and very poor in nutrients, leads to the disappearance of these microorganisms due to the toxic effect of bile acids. A multidimensional analysis of data in the form of principal component analysis indicated that lactic acid bacteria bind bile acids and show antagonistic effect on E. coli spp. bacteria. The growth in E. coli spp. population was accompanied by a decline in the population of Bifidobacterium spp. and Lactobacillus spp. with a simultaneous reduction in the concentration of bile acids. This is direct proof of acid binding ability of the tested lactic acid bacteria with respect to cholic acid, lithocholic acid and deoxycholic acid. This research demonstrated that the degree of fineness of wheat dietary fibre does not affect the sorption of bile acids and growth of some bacteria species; however, it has an impact on the profile of synthesized short-chained fatty acids. During the digestion of a very fine wheat fibre fraction (WF 90), an increase in the concentration of propionic and butyric acids, as compared with the wheat fiber fraction of larger particles - WF 500, was observed. Our study suggested that wheat fibre did not affect faecal bacteria growth, however, we observed binding of bile acids by Bifidobacterium spp. and Lactobacillus spp.

Hypothetical glycerol pathways of newly isolated strains capable of 1,3-propanediol production.

Study presented here demonstrates the ability of three newly isolated strains, obtained from environmental probes (manure, bottom sediment, and food waste) and identified as Clostridium bifermentans, Clostridium butyricum, and Hafnia alvei, to synthesize 1,3-propanediol (1,3-PD), organic acids (such as lactic, acetic, fumaric, succinic, and butyric acids), and ethanol from glycerol. The production of 1,3-PD as well as the glycerol pathways in C. bifermentans and H. alvei cells have not been investigated and described yet by others. Moreover, there is no data in the available literature on the products of glycerol utilization by H. alvei and there is only some incoherent data (mainly from the first half of the twentieth century) about the ability of C. bifermentans to carry out glycerol degradation. Additionally, this study presents complete hypothetical glycerol pathways and the basic fermentation kinetic parameters (such as yield and productivity) for both strains as well as for the newly isolated C. butyricum strain.

Effect of boiling in water of barley and buckwheat groats on the antioxidant properties and dietary fiber composition.

In recent years, there has been an ever-increasing interest in the research of polyphenols obtained from dietary sources, and their antioxidative properties. The purpose of this study was to determine the effect of boiling buckwheat and barley groats on the antioxidant properties and dietary fiber composition. Antioxidative properties were investigated using methyl linoleate model system, by assessing the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity and metal chelating activity. The results were compared with butylated hydroxytoluene (BHT). Raw barley and buckwheat groats extracts showed higher DPPH scavenging ability compared to boiled barley and buckwheat groats extracts. Raw barley groats extract exhibited higher antioxidant activity than boiled groats extract in the methyl linoleate emulsion. Higher chelating ability in relation to Fe (II) ions was observed for boiled groats extracts as compared to raw groats extracts. BHT showed small antiradical activity and metal chelating activity, while showing higher antioxidative activity in emulsion system. The analysis of groats extracts using HPLC method showed the presence of rutin, catechin, quercetin, gallic, p-hydroxybenzoic, p-coumaric, o-coumaric, vanillic, sinapic, and ferulic acids. Differences in the content of dietary fiber and its fractions were observed in the examined products. The highest total dietary fiber content was detected in boiled buckwheat groats, while the lowest - in boiled barley groats. The scientific achievements of this research could help consumers to choose those cereal products available on the market, such as barley and buckwheat groats, which are a rich source of antioxidative compounds and dietary fiber.

Conversion of glycerol to 1,3-propanediol by Citrobacter freundii and Hafnia alvei - newly isolated strains from the Enterobacteriaceae.

In this study, nearly 4000 bacterial strains from the family of Enterobacteriaceae isolated from different environments were screened for ability to convert glycerol to 1,3-propanediol (1,3-PD). The aim of the research was to isolate 1,3-PD producers from the natural environment, identify and characterize the best isolates. Three selective media were tested to usefulness in the isolation of bacteria from the family Enterobacteriaceae. Only, 28% of examined isolates could synthesize 1,3-PD from glycerol. 1,3-PD producing bacteria were identified by API 20E tests and 16S rRNA sequences to be Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter freundii and Hafnia alvei. It is the first time, when the fermentation glycerol to 1,3-PD by H. alvei was investigated. The selected strains (C. freundii AD119 and H. alvei AD27) were analyzed on a bioreactor scale under constant pH value 7.0 at temperature of 30°C and 37°C. After 40h in batch fermentation, H. alvei AD27 produced 11.3g/L of 1,3-PD at 37°C. For C. freundii AD119, the best results were obtained at temperature of 30°C. After 24h of fermentation, the 1,3-PD concentration reached above 23 g/L of 1,3-PD.

Denitrification of industrial wastewater: Influence of glycerol addition on metabolic activity and community shifts in a microbial consortium.

The wastewater originating from explosives manufacturing plants are characterized by a high concentration of nitrates (3200mgNL(-1)), sulfates (1470mgL(-1)) and low pH (1.5) as well as the presence of organic compounds, such as nitroglycerin (1.9mgL(-1)) and nitroglycol (4.8mgL(-1)). The application of glycerol (C/N=3) at such a high concentration enabled complete removal of nitrates and did not cause the anaerobic glycerol metabolic pathway of the DNC4 consortium to activate, as confirmed by the low concentrations of 1,3-propanediol (0.16gL(-1)) and acetic acid (0.11gL(-1)) in the wastewater. Increasing the glycerol content (C/N=5) contributed to a notable increase in the concentration of both compounds: 1.12gL(-1) for acetic acid and 1.82 for 1,3-PD (1,3-propanediol). The nitrate reduction rate was at 44mgNg(-1) biomass d(-1). In order to assess the metabolic activity of the microorganisms, a method to determine the redox potential was employed. It was established, that the microorganisms can be divided into four groups, based on the determined denitrification efficiency and zero-order nitrate removal constants. The first group, involving Pseudomonas putida and Pseudomonas stutzeri, accounts for microorganisms capable of the most rapid denitrification, the second involves rapid denitrifying microbes (Citrobacter freundi and Pseudomonas alcaligenes), the third group are microorganisms exhibiting moderate denitrification ability: Achrobactrum xylosoxidans, Ochrobactrum intermedium and Stenotrophomonas maltophila, while the last group consists of slow denitrifying bacteria: Rodococcus rubber and Sphignobacterium multivorum.

Biological Denitrification of High Nitrate Processing Wastewaters from Explosives Production Plant.

Wastewater samples originating from an explosives production plant (3,000 mg N l(-1) nitrate, 4.8 mg l(-1) nitroglycerin, 1.9 mg l(-1) nitroglycol and 1,200 mg l(-1) chemical oxygen demand) were subjected to biological purification. An attempt to completely remove nitrate and to decrease the chemical oxygen demand was carried out under anaerobic conditions. A soil isolated microbial consortium capable of biodegrading various organic compounds and reduce nitrate to atmospheric nitrogen under anaerobic conditions was used. Complete removal of nitrates with simultaneous elimination of nitroglycerin and ethylene glycol dinitrate (nitroglycol) was achieved as a result of the conducted research. Specific nitrate reduction rate was estimated at 12.3 mg N g(-1) VSS h(-1). Toxicity of wastewater samples during the denitrification process was studied by measuring the activity of dehydrogenases in the activated sludge. Mutagenicity was determined by employing the Ames test. The maximum mutagenic activity did not exceed 0.5. The obtained results suggest that the studied wastewater samples did not exhibit mutagenic properties.

Application of fast liquid chromatography for antioxidants analysis.

BACKGROUND: An intensive development of the Fast Liquid Chromatography (FLC) has been recently observed. It makes possible to reduce time analysis and improve resolution as well as sensitivity. The aim of this study was to separate the chosen antioxidants optimization using the FLC method. MATERIAL AND METHODS: The three various procedures for antioxidants analysis were compared. Mobile phases containing aqueous solution of formic acid, acetic acid, acetonitrile, and methanol were tested. Limit of detection (LOD), limit of quantification (LOQ), linearity and repeatability of each procedures were determined. RESULTS: Developed procedure enabled to separate all analytes and allowed to get low LOD levels and good repeatability. This procedure was used for antioxidants analysis in buckwheat and buckwheat products. CONCLUSION: Fast Liquid Chromatography allows to reduce time analysis and obtain good validation parameters.

Biological denitrification of brine: the effect of compatible solutes on enzyme activities and fatty acid degradation.

The effect of the addition of compatible solutes (ectoine and trehalose) on the denitrification process of saline wastewater was studied. In saline wastewater, it was observed that the initial concentration of nitrates was 500 mg N l⁻¹. A fatty substance isolated from oiled bleaching earth (waste of vegetable oil refining process) was used as a source of carbon.The consortium, which was responsible for the denitrification process originated from the wastewater of the vegetable oil industry. The consortium of microorganisms was identified by the use of restriction fragment length polymorphism of 16S rRNA gene amplicons and sequencing techniques. It was noted that ectoine affects significantly the activity of lipase and nitrate reductase, and resulted in faster denitrification compared to saline wastewater with the addition of trehalose or control saline wastewater (without compatible solutes). It was observed that relative enzyme activities of lipase and nitrate reductase increased by 32 and 35%, respectively, in the presence of 1 mM ectoine. This resulted in an increase in specific nitrate reduction rate in the presence of 1 mM ectoine to 5.7 mg N g⁻¹ VSS h⁻¹, which was higher than in the absence of ectoine (3.2 mg N g⁻¹ VSS h⁻¹). The addition of trehalose did not have an effect on nitrate removals. Moreover, it was found that trehalose was used up completely by bacteria as a source of carbon in the denitrification process. The fatty acids were biodegraded by 74% in the presence of 1 mM ectoine.

Vitamin B12 production from crude glycerol by Propionibacterium freudenreichii ssp. shermanii: optimization of medium composition through statistical experimental designs.

A two-step statistical experimental design was employed to optimize the medium for vitamin B(12) production from crude glycerol by Propionibacterium freudenreichii ssp. shermanii. In the first step, using Plackett-Burman design, five of 13 tested medium components (calcium pantothenate, NaH(2)PO(4)·2H(2)O, casein hydrolysate, glycerol and FeSO(4)·7H(2)O) were identified as factors having significant influence on vitamin production. In the second step, a central composite design was used to optimize levels of medium components selected in the first step. Valid statistical models describing the influence of significant factors on vitamin B(12) production were established for each optimization phase. The optimized medium provided a 93% increase in final vitamin concentration compared to the original medium.