Current challenges of microalgae applications: exploiting the potential of non-conventional microalgae species.

The intensified attention to health, the growth of an elderly population, the changing lifestyles, and the medical discoveries have increased demand for natural and nutrient-rich foods, shaping the popularity of microalgae products. Microalgae thanks to their metabolic versatility represent a promising solution for a 'green' economy, exploiting non-arable land, non-potable water, capturing carbon dioxide (CO2) and solar energy. The interest in microalgae is justified by their high content of bioactive molecules, such as amino acids, peptides, proteins, carbohydrates, polysaccharides, polyunsaturated fatty acids (as ω-3 fatty acids), pigments (as ß-carotene, astaxanthin, fucoxanthin, phycocyanin, zeaxanthin and lutein), or mineral elements. Such molecules are of interest for human and animal nutrition, cosmetic and biofuel production, for which microalgae are potential renewable sources. Microalgae, also, represent effective biological systems for treating a variety of wastewaters and can be used as a CO2 mitigation approach, helping to combat greenhouse gases and global warming emergencies. Recently a growing interest has focused on extremophilic microalgae species, which are easier to cultivate axenically and represent good candidates for open pond cultivation. In some cases, the cultivation and/or harvesting systems are still immature, but novel techniques appear as promising solutions to overcome such barriers. This review provides an overview on the actual microalgae cultivation systems and the current state of their biotechnological applications to obtain high value compounds or ingredients. Moreover, potential and future research opportunities for environment, human and animal benefits are pointed out. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effect of microbial fermentation on functional traits and volatiloma profile of pâté olive cake.

In this study, the pâté olive cake (POC), a by-product of the olive oil industry, was subjected to fermentation in a bioreactor using three microbial strains, Lactiplantibacillus plantarum, Wickerhamomyces anomalus and Candida boidinii, previously isolated from fermented table olive brines. Chemical, microbiological and molecular analyses were carried out at the beginning and at the end of fermentation. The lowest pH value (4.09) was reached after 10 days in sample inoculated with C. boidinii. Microbiological analyses exhibited the dominance of yeasts throughout the whole process (from 5.5 to 7.80 Log10 CFU/g), as confirmed by PCR-DGGE analysis. The microbial cultures affected both phenolic and volatile organic compound profiles. Moreover, the POC samples treated with different microbial strains were investigated for biological assays. The sample fermented with W. anomalus showed the greatest diffusion speed of transepithelial transport through Caco-2 cell, the highest inhibitory activity towards the tested cyclooxygenases and the highest antioxidant activity.

Inside Current Winemaking Challenges: Exploiting the Potential of Conventional and Unconventional Yeasts.

Wine represents a complex matrix in which microbial interactions can strongly impact the quality of the final product. Numerous studies have focused on optimizing microbial approaches for addressing new challenges to enhance quality, typicity, and food safety. However, few studies have investigated yeasts of different genera as resources for obtaining wines with new, specific traits. Currently, based on the continuous changes in consumer demand, yeast selection within conventional Saccharomyces cerevisiae and unconventional non-Saccharomyces yeasts represents a suitable opportunity. Wine fermentation driven by indigenous yeasts, in the various stages, has achieved promising results in producing wines with desired characteristics, such as a reduced content of ethanol, SO2, and toxins, as well as an increased aromatic complexity. Therefore, the increasing interest in organic, biodynamic, natural, or clean wine represents a new challenge for the wine sector. This review aims at exploring the main features of different oenological yeasts to obtain wines reflecting the needs of current consumers in a sustainability context, providing an overview, and pointing out the role of microorganisms as valuable sources and biological approaches to explore potential and future research opportunities.

Protective Effect of Treated Olive Mill Wastewater on Target Bacteria and Mitochondrial Voltage-Dependent Anion-Selective Channel 1.

Olive mill wastewater, a by-product of the olive oil industry, represents an important resource, rich in bioactive compounds with antioxidant activity. In this study, two strategies to concentrate the bioactive components were used: the tangential membrane filtration (ultrafiltration and reverse osmosis) and the selective resin extraction. The concentrates were evaluated for physico-chemical characteristics and antioxidant activity. Furthermore, the antimicrobial activity and the effect on the mitochondrial voltage-dependent anion selective channel 1 were evaluated. The chemical results highlighted that the highest concentration of hydroxytyrosol (as 7204 mg/L) was revealed in the sample obtained by inverse osmosis while the highest concentration of oleuropein (10005 mg/L) was detected in the sample obtained by resin extraction. The latter sample exhibited the highest antimicrobial effects against Listeria monocytogenes, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. Both samples exhibited a high impact on the electrophysiological parameters of VDAC1 activity. These results showed that both valorization techniques, which can be reproduced at industrial scale, provided phenolic concentrates with antioxidant and antimicrobial activity useful for different future perspectives.

Almond [Prunus dulcis (Mill.) DA Webb] Processing Residual Hull as a New Source of Bioactive Compounds: Phytochemical Composition, Radical Scavenging and Antimicrobial Activities of Extracts from Italian Cultivars ('Tuono', 'Pizzuta', 'Romana').

In this study we developed a new extract, by the use of conventional solid-solvent extraction and a food-grade hydroalcoholic solvent, rich in phenolic and triterpenoid components from almon hull to be employed as functional ingredient in food, pharma and cosmetic sectors. Two autochthonous Sicilian cultivars ('Pizzuta' and 'Romana') and an Apulian modern cultivar ('Tuono') have been tested for the production of the extract. Results showed that the two Sicilian varieties, and in particular the 'Romana' one, present the best characteristics to obtain extracts rich in triterpenoids and hydroxycinnamic acids, useful for the production of nutraceutical supplements. About triterpenoids, the performance of the hydroalcoholic extraction process allowed to never go below 46% of recovery for 'Pizzuta' samples, with significantly higher percentages of recovery for 'Tuono' and 'Romana' extracts (62.61% and 73.13%, respectively) while hydroxycinnamic acids were recovered at higher recovery rate (84%, 89% and 88% for 'Pizzuta', 'Romana' and 'Tuono' extracts, respectively). Invitro antioxidant and antimicrobial activities exerted by the extracts showed promising results with P. aeruginosa being the most affected strain, inhibited up to the 1/8 dilution with 'Romana' extract. All the three tested extracts exerted an antimicrobial action up to 1/4 dilutions but 'Romana' and 'Pizzuta' extracts always showed the greatest efficacy.

Olive Mill Wastewater Fermented with Microbial Pools as a New Potential Functional Beverage.

Olive mill wastewater (OMWW) represents a by-product but also a source of biologically active compounds, and their recycling is a relevant strategy to recover income and to reduce environmental impact. The objective of the present study was to obtain a new functional beverage with a health-promoting effect starting from OMWW. Fresh OMWW were pre-treated through filtration and/or microfiltration and subjected to fermentation using strains belonging to Lactiplantibacillus plantarum, Candida boidinii and Wickerhamomyces anomalus. During fermentation, phenolic content and hydroxytyrosol were monitored. Moreover, the biological assay of microfiltered fermented OMWW was detected versus tumor cell lines and as anti-inflammatory activity. The results showed that in microfiltered OMWW, fermentation was successfully conducted, with the lowest pH values reached after 21 days. In addition, in all fermented samples, an increase in phenol and organic acid contents was detected. Particularly, in samples fermented with L. plantarum and C. boidinii in single and combined cultures, the concentration of hydroxytyrosol reached values of 925.6, 902.5 and 903.5 mg/L, respectively. Moreover, biological assays highlighted that fermentation determines an increase in the antioxidant and anti-inflammatory activity of OMWW. Lastly, an increment in the active permeability on Caco-2 cell line was also revealed. In conclusion, results of the present study confirmed that the process applied here represents an effective strategy to achieve a new functional beverage.

Multi-Functional Potential of Lactic Acid Bacteria Strains and Antimicrobial Effects in Minimally Processed Pomegranate (Punica granatum L. cv Jolly Red) Arils.

This study aimed to evaluate the antimicrobial activity of both cells, and cell-free supernatants (CFS) of 7 selected lactic acid bacteria (LAB) strains belonging to Limosilactobacillus fermentum (4 strains), Lacticaseibacillus paracasei (1 strain), Lacticaseibacillus rhamnosus (1 strain), and Enterococcus faecium (1 strain) species, against Listeria monocytogenes, Escherichia coli, Salmonella Typhimurium, Pseudomonas aeruginosa and Staphylococcus aureus, by both the agar-well diffusion and co-culture methods. In addition, probiotic and safety traits were also detected. Great variability was detected on antimicrobial effects, whereas all tested strains were found sensitive to most of the tested antibiotics, and without any DNase, gelatinase, or hemolytic activity. Moreover, strains showed excellent survival in acidic conditions and exhibited tolerance to pepsin and bile salts. Based on the in vitro results, the CFSs of two selected L. fermentum strains were applied, in a mixed solution, as bio-preservative into minimally processed pomegranate arils, inoculated with a cocktail of L. monocytogenes and E. coli. Samples, packaged in an ordinary atmosphere, were analyzed during refrigerated storage, for up to 12 days, for physicochemical (as weight loss, texture, color, pH, total soluble solids and organic acid content) and for microbiological traits. Results revealed the effectiveness of CFS, up to 12 days, in reducing weight loss and microbial growth, without any significant effect on texture, total soluble solid content and color, found comparable to the acid citric treatment, highlighting the multi-functional potential of selected probiotic strains.

Phenols recovered from olive mill wastewater as natural booster to fortify blood orange juice.

In the present study, a tangential membrane filtration system was applied to recover phenols from olive mill wastewater. The obtained concentrates were characterised for physico-chemical traits, antioxidant activity and antimicrobial effects. Results indicated that the highest concentration of hydroxytyrosol (7203.7 mg/L) was detected in the concentrate obtained by reverse osmosis, which also showed the highest antioxidant and antimicrobial activity. Moreover, the same concentrate was added, at different ratio, up to 4:250 v/v, into a commercial blood orange juice. The fortified juice with the addition of the concentrate, up to 2:250 v/v ratio, did not show off-flavour and off-odour compared to the control. Furthermore, after 60 days of refrigerated storage, the fortified juice exhibited a hydroxytyrosol content still complying with the daily intake recommended by EFSA health claim. The obtained results can be industrially useful in producing orange juice added with a natural antioxidant concentrate as a 'clean label' ingredient.

Olive Pomace and Pâté Olive Cake as Suitable Ingredients for Food and Feed.

Olive oil extraction generates several by-products that represent an environmental issue, mainly for Mediterranean countries where olive oil is mostly produced. These by-products represent an ecological issue for their phenolic components, such as oleuropein, hydroxytyrosol, and tyrosol. However, olive oil by-products can be treated and properly exploited in different fields for their health-promoting properties, and they represent great potential for the food and beverage, cosmetic, and pharmaceutical industries. Furthermore, recovery and treatment processes can contribute to efficient waste management, which can enhance the sustainability of the olive oil industry, and in turn, lead to relevant economic benefits. The solid waste, i.e., olive pomace, could be considered to be a suitable matrix or primary resource of molecules with high added value due to their high phenolic content. Olive pomace, at different moisture contents, is the main by-product obtained from two- or three-phase extraction systems. A commonly used centrifugal extraction system, i.e., a multiphase decanter (DMF), does not require the addition of water and can generate a new by-product called pâté or olive pomace cake, consisting of moist pulp that is rich in phenols, in particular, secoiridoids, without any trace of kernel. Although several reviews have been published on olive wastes, only a few reviews have specifically focused on the solid by-products. Therefore, the aim of the present review is to provide a comprehensive overview on the current valorization of the main solid olive oil by-products, in particular, olive pomace or pâté olive cake, highlighting their use in different fields, including human nutrition.

Prebiotic effects of citrus pectic oligosaccharides.

Citrus peel wastes, one of the major by-products of the agri-food industry, are a source of value-added compounds. In this work, a commercial pectin (PEC1) and one obtained by direct extraction from citrus fibre (PEC2), were hydrolysed and ultrafiltered at different cut-off (100, 50 and 30 kDa) and then tested in broth with four different probiotic strains: Lacticaseibacillus paracasei, Limosilactobacillus fermentum, Lacticaseibacillus rhamnosus and Bifidobacterium animalis subsp. paracasei. In addition, the probiotic strains selected for their growth rate with the fractions of the two pectins were tested for tolerance to different pH values and bile salts. This study demonstrated that POS2 (product from the enzymatic hydrolysis by PEC2) showed greater prebiotic activity than POS1 (product from the enzymatic hydrolysis by PEC1). The results confirmed that citrus pectic oligosaccharides could be used as emerging prebiotics with improved properties due to their antimicrobial and modulating-microbiota ability.

Microbial Application to Improve Olive Mill Wastewater Phenolic Extracts.

Olive mill wastewater (OMW) contains valuable and interesting bioactive compounds, among which is hydroxytyrosol, which is characterized by a remarkable antioxidant activity. Due to the health claims related to olive polyphenols, the aim of this study was to obtain an extract from OMW with an increased level of hydroxytyrosol by means of microbial enzymatic activity. For this purpose, four commercial adsorbent resins were selected and tested. The beta-glucosidase and esterase activity of strains of Wickerhamomyces anomalus, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae were also investigated and compared to those of a commercial enzyme and an Aspergillus niger strain. The W. anomalus strain showed the best enzymatic performances. The SP207 resin showed the best efficiency in selective recovery of hydroxytyrosol, tyrosol, oleuropein, and total phenols. The bioconversion test of the OMW extract was assessed by using both culture broths and pellets of the tested strains. The results demonstrated that the pellets of W. anomalus and L. plantarum were the most effective in hydroxytyrosol increasing in phenolic extract. The interesting results suggest the possibility to study new formulations of OMW phenolic extracts with multifunctional microorganisms.

Metagenetic and Volatilomic Approaches to Elucidate the Effect of Lactiplantibacillus plantarum Starter Cultures on Sicilian Table Olives.

The present study aimed to evaluate the effect of selected Lactiplantibacillus plantarum strains on both microbiota composition and volatile organic compound profile of Sicilian table olives. Two mixed cultures, named O1 and O2, were set up for pilot-plan scale fermentations at 5% of NaCl. Uninoculated table olives at 5 and 8% (C5 and C8) of salt were used as control. The fermentation process was monitored until 80 days through a dual approach, which included both classical microbiological and 16S amplicon-based sequencing and volatilomics analyses. Compared with control samples (C5 and C8), experimental samples, inoculated with starter cultures (O1 and O2), exhibited a faster acidification with a more pronounced drop in pH. Metagenetics data revealed significant differences of microbiota composition among samples, highlighting the dominance of lactobacilli in both experimental samples; a high occurrence of Enterobacter genus only in control samples with 5% of NaCl; and the presence of Bacteroides, Faecalibacterium, Klebsiella, and Raoultella genera only in control samples with 8% of NaCl. Furthermore, microbiota composition dynamics, through the fermentation process, significantly affected the volatile organic compounds of the final products, whereas no compounds involved in off-odors metabolites were detected in all samples investigated. In conclusion, the addition of the proposed starter cultures and the use of low concentrations of sodium chloride positively affected the microbiota and volatile organic compounds, ensuring the microbiological safety and the pleasant flavors of the final product.

Effects of Different Stress Parameters on Growth and on Oleuropein-Degrading Abilities of Lactiplantibacillus plantarum Strains Selected as Tailored Starter Cultures for Naturally Table Olives.

The use of ß-glucosidase positive strains, as tailored-starter cultures for table olives fermentation, is a useful biotechnological tool applied to accelerate the debittering process. Nowadays, strains belonging to Lactiplantibacillus plantarum species are selected for their high versatility and tolerance to stress conditions. The present study investigated the effect of different stress factors (pH, temperature and NaCl) on growth and on oleuropein-degrading abilities of selected L. plantarum strains. In addition, the presence of the beta-glucosidase gene was investigated by applying a PCR based approach. Results revealed that, overall, the performances of the tested strains appeared to be robust toward the different stressors. However, the temperature of 16 °C significantly affected the growth performance of the strains both singularly and in combination with other stressing factors since it prolongs the latency phase and reduces the maximum growth rate of strains. Similarly, the oleuropein degradation was mainly affected by the low temperature, especially in presence of low salt content. Despite all strains displayed the ability to reduce the oleuropein content, the beta-glucosidase gene was detected in five out of the nine selected strains, demonstrating that the ability to hydrolyze the oleuropein is not closely related to the presence of beta-glucosidase. Data of the present study suggest that is extremely important to test the technological performances of strains at process conditions in order to achieve a good selection of tailored starter cultures for table olives.

Pilot study to assess isoflavone intake in middle-aged italian subjects.

Knowledge of isoflavone (IF) intake in Western populations is scarce, primarily because data about the content of these compounds in non-soy derived foods are incomplete or unavailable. The aims of this study were 1) to enrich the data available in literature about the IF content in traditional Italian foods, 2) to estimate daidzein (D) and genistein (G) intake in an Italian population sample. Eighteen Italian foods have been selected and analysed for IF content by GC-MS; the assessment of IF intake was performed in sixty healthy middle-aged Italian subjects after investigation of their dietary habits by food frequency questionnaire (FFQ). The mean IF intake was 171 ± 261 µg/die (26-1415 µg/die). The mean G intake was greater than D (98 ± 131 µg/die vs 76 ± 131 µg/die). As expected, soy products, even though poorly consumed (27%), resulted the main contributor to IF intake (IF intake was 473.4 ± 440 µg/die vs 75 ± 38 µg/die in soy consumers and non soy consumers respectively p<0.001). Among Mediterranean foods, the main contributor resulted fresh bread that is widely consumed (97%). The percentage contribution of the cereal group to mean IF intake was 91%; the legume, fruit and vegetables groups brought a low contribution (3%, 2% and 4% respectively). The total daily IF intake found was low and probably not sufficient to produce biological effects. However more studies are necessary to investigate whether low exposure to IF for a long time could have positive effects on human health.

Effectiveness of moderate green tea consumption on antioxidative status and plasma lipid profile in humans.

The antioxidant activity of green tea (GT) has been extensively studied; however, the results obtained from dietary intervention studies are controversial. In the present study we investigated the effect of the addition of two cups of GT (containing approximately 250 mg of total catechins) to a controlled diet in a group of healthy volunteers with respect to a group following the same controlled diet but not consuming GT. Antioxidant status and lipid profile in plasma, the resistance from oxidative damage to lipid and DNA, and the activity of glutathione peroxidase (GPX) in isolated lymphocytes were measured at the beginning and the end of the trial. After 42 days, consumption of GT caused a significant increase in plasma total antioxidant activity [from 1.79 to 1.98 micromol Trolox equivalent (TE)/ml, P<.001], significant decreases in plasma peroxides level (from 412 to 288 Carr U, P<.05) and induced DNA oxidative damage in lymphocytes (from 14.2% to 10.1% of DNA in tail, P<.05), a moderate although significant decrease in LDL cholesterol (from 119.9 to 106.6 mg/dL, P<.05) with respect to control. The present study suggests the ability of GT, consumed within a balanced controlled diet, to improve overall the antioxidative status and to protect against oxidative damage in humans.

Selection of a WEHI-3B leukemia cell subclone resistant to inhibition by cholera toxin.

The studies on the inhibitory effect exerted by Cholera Toxin (CT) on cell growth and proliferation indicate a remarkable heterogeneity of cell response suggesting that the inhibition represents the final event of many different ways or mechanisms. After CT binding, cAMP accumulation may not occur (as in L1210 leukemia cells) or, when occurring (as in SR-4987 stromal cells), may not be coupled with the antiproliferative effect of CT. In WEHI-3B cells CT binds a Gal-GalNac-GM1b receptor and the anticlonogenic effect of CT seems correlated with cAMP accumulation. To demonstrate the central role of cAMP in WEHI-3B cells, starting from the sensitive cell strain we selected and established a clone of WEHI-3B resistant to CT. This revertant clone (WEHI-3B/CT/REV) is currently cultured in the absence of CT and in the proliferation assay shows a dramatic resistance (>46,000 than the parental cells). Stimulation ofWEHI-3B/CT/REV cells by cholera toxin failed to enhance cAMP and the ganglioside-CT binding studied on Thin Layer Chromatography (TLC) blots showed that the resistant cells lost the spot correspondent to the migration of Gal-GalNac-GM1b ganglioside. Both the lines respond at the same level to the adenylate cyclase stimulation by forskolin and the incorporation of GM1a did not decrease the resistance of WEHl-3B/CT/REV. These data confirm that Gal-GalNac-GM1b is the most important functional receptor for CT in WEHI-3B cells able to transduce the signal by enhancing cAMP which in turn inhibits cell proliferation (probably by cAMP dependent protein kinase activation). Our study describes the first cell line resistant to CT originated from a susceptible parental strain and provides a new interesting cell model for studying the cAMP dependent mechanisms involved in cell growth regulation.

Secretion of basic fibroblast growth factor (FGF-2) by WEHI-3B myelomonocytic leukemia cells.

In order to investigate the role of Fibroblast Growth Factors in hematopoietic cells, we studied the expression of FGF-1, FGF-2, FGF-3, FGF-4, FGF-5 and FGF-6 mRNAs both in murine myelomonocytic leukemia WEHI-3B and in a murine stromal cell line SR-4987. Secretion of FGF-2 in the cell culture supernatant was also studied. Expression of mRNA encoding for the above-mentioned FGFs was analyzed by RT-PCR. The production of FGF-2 in the conditioned media of WEHI-3B and SR-4987 cell cultures was evaluated by techniques of affinity chromatography, chromatofocusing and immunoblotting. The biological activity of FGF-2 was checked on SR-4987 cells by a agar clonogenic assay. In both cell lines mRNA was found encoding for FGF-1, FGF-2 and FGF-6 and WEHI-3B cells express also mRNA for FGF-3 (int-2) and FGF-4 (K-FGF/hst). Furthermore, supernatant from WEHI-3B cells was found to stimulate dramatically the agar clonogenicity of SR-4987 cells which have a very poor basal capacity for growth in agar. The clonogenic activity of WEHI-3B conditioned medium is due to FGF-2 secreted into cell culture supernatant whereas SR-4987 cells, although express FGF-2 mRNA, do not seem able to secrete this factor. The expression in myeloid leukemia cells of oncogene-related factors such as FGF-3, FGF-4 and FGF-6 together with the secretion of FGF-2 able to support a positive regulation of bone marrow stromal cells function suggest that FGFs may have an important role in sustaining the leukemogenic process and related disorders.