Supercritical CO2 Processing of a Functional Beverage Containing Apple Juice and Aqueous Extract of Pfaffia glomerata Roots: Fructooligosaccharides Chemical Stability after Non-Thermal and Thermal Treatments.

The effects of supercritical CO2 processing on the chemical stability of fructooligosaccharides (FOS) and other functional and nutritional compounds were evaluated employing non-thermal and thermal approaches. Apple juice was enriched with Pfaffia glomerata roots aqueous extract due to its high content of short-chain FOS and then subjected to different levels of temperature (40 and 60 °C), pressure (8 and 21 MPa), and CO2 volume ratio (20 and 50%). The percentage of CO2 volume was evaluated concerning the total volume of the high-pressure reactor. Also, the functional beverage was thermally treated at 105 °C for 10 min. Physicochemical properties (pH and soluble solid content), beta-ecdysone, sugars (glucose, fructose, and sucrose), and FOS (1-kestose, nystose, and fructofuranosylnystose) content were determined. The pH and soluble solid content did not modify after all treatments. The pressure and CO2 volume ratio did not influence the FOS content and their chemical profile, however, the temperature increase from 40 to 60 °C increased the nystose and fructofuranosylnystose content. High-temperature thermal processing favored the hydrolysis of 1-kestose and reduced the sucrose content. Regarding beta-ecdysone, its content remained constant after all stabilization treatments demonstrating thus its high chemical stability. Our results demonstrated that supercritical CO2 technology is a promising technique for the stabilization of FOS-rich beverages since the molecular structures of these fructans were preserved, thus maintaining their prebiotic functionality.

Effect of CO2 Flow Rate on the Extraction of Astaxanthin and Fatty Acids from Haematococcus pluvialis Using Supercritical Fluid Technology.

Haematococcus pluvialis is the largest producer of natural astaxanthin in the world. Astaxanthin is a bioactive compound used in food, feed, nutraceutics, and cosmetics. In this study, astaxanthin extraction from H. pluvialis by supercritical fluid extraction was evaluated. The effects of temperature (40 and 50 °C), pressure (40 and 50 MPa), and CO2 flow rate (2 and 4 L/min) were investigated. The results showed that the highest astaxanthin recovery was obtained at 50 °C/50 MPa and the CO2 flow rates evaluated had no significant effect. It was possible to achieve astaxanthin recoveries of 95% after 175 min for a CO2 flow rate of 2 L/min, and 95 min for CO2 flow rate of 4 L/min. The ω-6/ω-3 ratios obtained were similar in all conditions, reaching 0.87, demonstrating that the extracts from H. pluvialis by SFE are rich in unsaturated fatty acids (UFA) which increases their positive effects when used as a functional ingredient in food.

Effect of incorporation of antioxidants on the chemical, rheological, and sensory properties of probiotic petit suisse cheese.

This work investigated the effect of the addition of different antioxidants (ascorbic acid, glucose oxidase, cysteine, and jabuticaba extract) on the rheological and sensorial properties of the probiotic petit suisse cheese. Absence of influence of the antioxidants at the physico-chemical characteristics of the petit suisse cheese was observed. Overall, the petit suisse cheeses presented weak gel characteristics and behaved as pseudoplastic material, except for control. All treatments exhibited a thixotropic non-Newtonian behavior; however, higher hysteresis area was obtained for control sample, which indicates that antioxidants incorporated to petit suisse had a protective effect on the typical thixotropic behavior of the Quark gel. The commercial sample presented higher scores for all aspects by consumers, whereas the probiotic petit suisse samples presented opposite behavior. Projective mapping was able to generate a vocabulary where the sample containing jabuticaba skin extract obtained by supercritical extraction was characterized by the panelists as presenting grape flavor and purple color.

Subcritical and supercritical technology for the production of second generation bioethanol.

There is increased interest in reducing our reliance on fossil fuels and increasing the share of renewable raw materials in our energy supply chain due to environmental and economic concerns. Ethanol is emerging as a potential alternative to liquid fuels due to its eco-friendly characteristics and relatively low production costs. As ethanol is currently produced from commodities also used for human and animal consumption, there is an urgent need of identifying renewable raw materials that do not pose a competitive problem. Lignocellulosic agricultural residues are an ideal choice since they can be effectively hydrolyzed to fermentable sugars and integrated in the context of a biorefinery without competing with the food supply chain. However, the conventional hydrolysis methods still have major issues that need to be addressed. These issues are related to the processing rate and generation of fermentation inhibitors, which can compromise the quality of the product and the cost of the process. As the knowledge of the processes taking place during hydrolysis of agricultural residues is increasing, new techniques are being exploited to overcome these drawbacks. This review gives an overview of the state-of-the-art of hydrolysis with subcritical and supercritical water in the context of reusing agricultural residues for the production of suitable substrates to be processed during the fermentative production of bioethanol. Presently, subcritical and/or supercritical water hydrolysis has been found to yield low sugar contents mainly due to concurrent competing degradation of sugars during the hydrothermal processes. In this line of thinking, the present review also revisits the recent applications and advances to provide an insight of future research trends to optimize on the subcritical and supercritical process kinetics.

In vitro susceptibility of Brazilian Pythium insidiosum isolates to essential oils of some Lamiaceae family species.

The present study aimed to evaluate the in vitro antimicrobial action of Origanum vulgare, Origanum majorana, Mentha piperita and Rosmarinus officinalis on Pythium insidiosum oomycete zoospores. The antimicrobial activity evaluation was performed by the broth microdilution method according to CSLI M38-A2 documentation adapted to phytopharmaceuticals. Twenty-two P. insidiosum isolates were evaluated, and the minimum inhibitory concentration was determined at 100% growth inhibition. All P. insidiosum isolates evaluated showed a minimum inhibitory concentration ranging from 0.05 to 1.75 mg/mL when O. vulgare oil was used and from 0.11 to 3.5 mg/mL for O. majorana, M. piperita and R. officinalis oils. The results obtained indicate that the essential oils tested showed antimicrobial activity on P. insidiosum, with O. vulgare essential oil showing the best performance. These findings emphasize the potential use of plant essential oils as control agents in P. insidiosum infections; further research, however, is needed so as the in vivo activity of these oils can also be evaluated.

In vitro susceptibility of zoospores and hyphae of Pythium insidiosum to antifungals.

OBJECTIVES: The purpose of this study was to compare the in vitro susceptibilities of 22 Brazilian isolates of Pythium insidiosum to antifungals using a standardized inoculum of zoospores and a proposed novel inoculum prepared from cultured mycelia (hyphae) of P. insidiosum. METHODS: A zoospore suspension of P. insidiosum was obtained by the zoosporogenesis technique. The hyphal inoculum was prepared from a suspension of P. insidiosum mycelium. Susceptibility to each drug was evaluated using the CLSI M38-A2 method. RESULTS: Of the 88 MIC comparisons performed, 36 (41%) showed the same MIC value for the two inocula. The agreement (differences not greater than one dilution) between MICs obtained with both types of inocula was 39.8% (35/88). In other MIC comparisons analysed, 17 (19.3%) showed differences of two or three dilutions. CONCLUSIONS: We conclude that the use of hyphal inocula of P. insidiosum for in vitro susceptibility tests could be a suitable method for evaluating antimicrobial susceptibility, particularly when it is not possible to obtain a standardized zoospore inoculum.

Wild animals as sentinels of Paracoccidioides brasiliensis in the state of Rio Grande do Sul, Brazil.

Paracoccidioides brasiliensis, a dimorphic pathogenic fungus, causes the principal form of systemic mycosis in Brazil. The literature furnishes only limited data on the ecology of this fungus in the state of Rio Grande do Sul, the southernmost state of Brazil. The purpose of this study was to evaluate the prevalence of fungal infection in wild animals, using serological tests and using the animals as sentinels of the presence of P. brasiliensis in three specified mesoregions of Rio Grande do Sul. A total of 128 wild animals from the three mesoregions were included in the study. The serum samples were evaluated by immunodiffusion and the enzyme-linked immunosorbent assay (ELISA) technique to detect anti-gp43 antibodies from P. brasiliensis. Two conjugates were tested and compared with the ELISA technique. Although no positive samples were detected by immunodiffusion, 26 animals (20%), belonging to 13 distinct species, were found to be seropositive by the ELISA technique. The seropositive animals were from two mesoregions of the state. The results were similar according to the gender, age, and family of the animals, but differed significantly according to the conjugate used (p < 0.001), showing more sensitivity to protein A-peroxidase than to protein G-peroxidase. The finding that wild animals from the state of Rio Grande do Sul are exposed to P. brasiliensis suggests that the fungus can be found in this region despite the often-rigorous winters, which frequently include below-freezing temperatures.

Characterization of a commercial cellulase for hydrolysis of agroindustrial substrates.

This work is focused on the characterization of a commercial cellulase in terms of optimum pH and temperature, stability to pH and temperature and affinity of this enzyme to several substrates, determining the Michaelis-Menten parameters. Maximum activity of cellulase was obtained for the temperature range from 40 to 50 °C and pH from 5.2 to 5.5. Enzyme activity decreased only 15% after 150 h of reaction at temperatures between 30 and 50 °C. No loss of activity was observed at pH 5.0 and 5.5. The cellulase showed satisfactory results in the hydrolysis of agroindustrial substrates, since similar activity was verified on filter paper and other agroindustrial substrates.

Interactions of ß-carotene with WPI/Tween 80 mixture and oil phase: Effect on the behavior of O/W emulsions during in vitro digestion.

This study investigated the impact of adding ß-carotene on the structure of fresh O/W emulsions with different oil phase (sunflower oil-LCT or NEOBEE®1053-MCT) and emulsifiers (WPI, Tween 80 - T80 or WPI/T80 mixture). In this sense, the behavior of emulsions through the gastrointestinal tract, the stability and bioaccessibility of ß-carotene were also assessed. The ß-carotene reduced the interfacial tension of the LCT/MCT-water systems. The addition of ß-carotene promoted an increase of viscoelasticity of LCT/MCT-T80 (0.5%WPI/0.5%T80 and 1%T80 w/w) interfaces, but an increase of WPI content reduced the viscoelasticity of interfacial layers (LCT/MCT-1% WPI). These changes in the interface properties influenced the mean droplet size and ζ-potential of the fresh emulsions. LCT systems presented similar bioaccessibility/stability of ß-carotene. However, ß-carotene entrapped within protein-coated MCT droplets was more stable than within T80-MCT systems. Our results show that ß-carotene interacted with other ingredients of emulsions changing their properties and behavior under gastrointestinal tract as well as the stability/bioaccessibility of ß-carotene.

Can the essential oil of rosemary (Rosmarinus officinalis Linn.) protect rats infected with itraconazole-resistant Sporothrix brasiliensis from fungal spread?

INTRODUCTION: Itraconazole is the first-choice option to treat human and animal sporotrichosis. However, the emergence of itraconazole-resistant strains has encouraged research on new active antifungals. Among them, the essential oil of rosemary (Rosmarinus officinalis Linn., Lamiaceae) has shown antifungal activity in vitro. OBJECTIVE: Assessing, for the first time, the effectiveness of rosemary essential oil in vivo in experimental cutaneous sporotrichosis, as well as its chemical composition and action mode. METHODS: Itraconazole-resistant Sporothrix brasiliensis was inoculated in the left foot pad of 30 Wistar rats, which were randomized (n=10) for treatment with saline solution (control, CONT), itraconazole (ITRA, 10 mg/kg) and rosemary oil (ROSM, 250 mg/kg) for 30 days at an oral dose of 1 mL, daily. Clinical evolution, histopathology and fungal burden were investigated. GC-MS was used for chemical analysis; sorbitol protection and ergosterol effect were used to evaluate the action mechanism of rosemary oil. RESULTS: ROSM was the only group evolving to skin lesion remission, lack of edema and exudate, and mild-to-absent yeast cells. Rosemary oil delayed fungal spreading and protected systemic organs, mainly liver and spleen. The ROSM group presented lower fungal load than that observed for the CONT and ITRA groups (p<0.05). Antifungal action took place at complexation level after ergosterol application. Most compounds were 1,8-cineole/eucalyptol (47.91%), camphor (17.92%), and α-pinene (11.52%). CONCLUSIONS: These findings have evidenced that rosemary oil is a promising antifungal to treat sporotrichosis, since it protects systemic organs from fungal spread.

Low-frequency and high-power ultrasound-assisted production of natural blue colorant from the milk and unripe Genipa americana L.

This study presents the production of a novel natural blue colorant obtained from the cross-linking between milk proteins and genipin assisted by low-frequency and high-power ultrasound technology. Genipin was extracted from unripe Genipa americana L. using milk as a solvent. Also, milk colloidal system was used as a reaction medium and carrier for the blue color compounds. The effects of ultrasound nominal power (100, 200, 300, and 400 W) on the blue color formation kinetics in milk samples were evaluated at 2, 24, and 48 h of cold storage in relation to their free-genipin content and color parameters. In addition, Fourier transform infrared (FTIR) spectrum, droplet size distribution, microstructure, and kinetic stability of the blue colorant-loaded milk samples were assessed. Our results have demonstrated that the ultrasound technology was a promising and efficient technique to obtain blue colorant-loaded milk samples. One-step acoustic cavitation assisted the genipin extraction and its diffusion into the milk colloidal system favoring its cross-linking with milk proteins. Ultrasound process intensification by increasing the nominal power promoted higher genipin recovery resulting in bluer milk samples. However, the application of high temperatures associated with intensified acoustic cavitation processing favored the occurrence of non-enzymatic browning due to the formation of complex melanin substances from the Maillard reaction. Also, the blue milk samples were chemically stable since their functional groups were not modified after ultrasound processing. Likewise, all blue colorant-loaded milk samples were kinetically stable during their cold storage. Therefore, a novel natural blue colorant with high-potential application in food products like ice creams, dairy beverages, bakery products, and candies was produced.

High-intensity ultrasound energy density: How different modes of application influence the quality parameters of a dairy beverage.

This study evaluated the influence of the high-intensity ultrasound (HIUS) technology on the quality parameters of a model dairy beverage (chocolate whey beverage), operating under the same energy density (5000 J/mL), but applied at different ways. Two processes were performed varying nominal power and processing time: HIUS-A (160 W and 937 s), and HIUS-B (720 W and 208 s). Our objective was to understand how different modes of application of the same HIUS energy density could influence the microstructure, droplet size distribution, zeta potential, phase separation kinetic, color parameters and mineral profile of the chocolate whey beverage. The results demonstrated that the different modes of application of the same HIUS energy density directly influenced the final quality of the product, resulting in whey beverages with distinct physical and microstructural characteristics. The HIUS-B processing was characterized as a thermal processing, since the final processing temperature reached 71 °C, while the HIUS-A processing was a non-thermal process, reaching a final temperature of 34 °C. Moreover, HIUS-B process greatly reduced the droplet size and increased the lightness value in relation to the HIUS-A processing. Both treatments resulted in whey beverages with similar phase separation kinetics and were more stable than the untreated sample. The HIUS processes did not modify the mineral content profile. Overall, the study emphasizes the versatility of the HIUS technology, highlighting that the processing must not be based only on the applied energy density, since different powers and processing times produce dairy beverages with distinct characteristics.

Xylooligosaccharides chemical stability after high-intensity ultrasound processing of prebiotic orange juice.

The effects of the high-intensity ultrasound (HIUS) technology at the nominal powers of 300, 600, 900, and 1200 W were evaluated on the chemical stability of xylooligosaccharides (XOS) used to enrich orange juice. The ultrasound energy performance for each nominal power applied to the XOS-enriched orange juice was determined by calculating acoustic powers (W), HIUS intensity (W/cm2), and energy density (kJ/mL). Physicochemical properties (pH and soluble solid content), organic acid content (ascorbic, malic, and citric acids), total phenolic content (TPC), antioxidant activity by the FRAP (Ferric reducing ability of plasma) method, sugar (glucose, fructose, and sucrose), and XOS (xylobiose, xylotriose, xylotetraose, xylopentaose, and xylohexaose) content were determined. The pH and soluble solid content did not change after all HIUS treatments. The HIUS process severity was monitored by quantifying ascorbic acid content after the treatments. A significant linear decrease in the ascorbic acid content was observed in prebiotic orange juice with the HIUS process intensification by increasing nominal power. The malic acid and citric acid contents had similar behavior according to the HIUS process intensification. The nominal power increase from 300 to 600 W increased the concentration of both organic acids, however, the intensification up to 1200 W reduced their concentration in the functional beverage. The TPC and FRAP data corroborated with the results observed for the ascorbic acid content. However, the HIUS processing did not alter sugar and XOS contents. The XOS chromatographic profiles were not modified by the HIUS treatment and presented the same amount of all prebiotic compounds before and after the HIUS treatment. Overall, HIUS technology has been evaluated as a promising stabilization technique for prebiotic beverages enriched with XOS due to their high chemical stability to this emerging technology under severe process conditions.

Ultrasound stabilization of raw milk: Microbial and enzymatic inactivation, physicochemical properties and kinetic stability.

The aim of this study was to evaluate the effects of non-thermal and thermal high-intensity ultrasound (HIUS) treatment on the microbial and enzymatic inactivation, physicochemical properties, and kinetic stability of the raw milk by applying different energy densities (1, 3, 5, and 7 kJ/mL). Two HIUS treatments were evaluated based on different nominal powers, named HIUS-A and HIUS-B, using 100 W and 475 W, respectively. HIUS-A treatment was non-thermal processing while HIUS-B was a thermal treatment only for the energy densities of 5 and 7 kJ/mL since the final temperature was above 70 °C. The HIUS-B treatment showed to be more efficient. Log reductions up to 3.9 cycles of aerobic mesophilic heterotrophic bacteria (AMHB) were achieved. Significant reductions of the fat globule size, with diameters lower than 1 µm, better color parameters, and kinetic stability during the storage were observed. Also, HIUS-B treatment inactivated the alkaline phosphatase and lactoperoxidase. The HIUS-B treatment at 3 kJ/mL worked below 57 °C being considered a border temperature since it did not cause unwanted physicochemical effects. Furthermore, a microbial inactivation of 1.8 ± 0.1 log cycles of AMHB was observed. A proper inactivation of only the Alkaline phosphatase and a significant reduction of the fat globules sizes, which kept the milk kinetically stable during storage was achieved.

Extraction of bioactive compounds from defatted passion fruit bagasse (Passiflora edulis sp.) applying pressurized liquids assisted by ultrasound.

Passion fruit bagasse is a rich source of phenolic compounds, including piceatannol, a stilbene to which several biological activities are conferred. This work reports the application of pressurized liquid extraction (PLE) assisted by ultrasound (US) to intensify the extraction of phenolic compounds from defatted passion fruit bagasse (DPFB). PLE at different temperatures (65-75 °C) without and with different US powers (240-640 W) was performed to investigate the mechanism of the assisted process. The extracts were evaluated in terms of global, total phenolic (TP), piceatannol and total reducing sugar yields. The antioxidant capacity of the extracts was determined by FRAP and ORAC assays. PLE assisted by US increased the yields, resulting in 60% more TP and piceatannol. The observed yields suggest that the main mechanism driving PLE assisted by US from DPFB was the rise in temperature caused by the ultrasonic waves. Pearson coefficient revealed a strong correlation between antioxidant capacity and total phenolics and piceatannol yield. The three-line spline model was adequately fitted to the experimental curves, showing three extraction periods in which the recovery of TP and piceatannol was higher than 70% at the end of the falling extraction rate period. PLE assisted or not by US showed to be clean, efficient and green alternatives for the recovery of phenolic compounds. The findings of this work indicate that PLE assisted by US has a great potential to improve the extraction of bioactive compounds from natural products.

Impregnation of passion fruit bagasse extract in alginate aerogel microparticles.

Passion fruit bagasse extract (PFBE) is a rich source of polyphenols, including piceatannol. This work produced alginate (1, 2, 3 wt%) aerogel and investigated the impregnation of gallic acid (GA) and PFBE in alginate aerogel microparticles. The microparticles of ca. 100 µm in diameter were obtained by emulsion-gelation method, submitted to solvent exchange, wet impregnation (WI) and supercritical drying. Alginate aerogels derived from 1 wt% solution led to a higher GA loading and, therefore, this formulation was used to impregnate PFBE. The loading of PFBE, total phenolic, and piceatannol contents based on grams of raw aerogel were 0.62 g, 10.77 mg, and 741.85 µg, respectively, which means a loading efficiency of total phenolics and piceatannol of 47.1% and 34.7%. DSC analysis and X-ray diffraction showed that particles behave as amorphous materials and ORAC assay revealed that impregnated aerogel microparticles presented antioxidant capacity. Alginate aerogel microparticles presented as an appropriated material for drug loading, whereas WI and supercritical drying demonstrated to be useful techniques to load PBBE in aerogels.

Effects of supercritical carbon dioxide and thermal treatment on the inulin chemical stability and functional properties of prebiotic-enriched apple juice.

Inulin-enriched apple juice was subjected to supercritical CO2 processing under different pressure levels (10, 15, and 20 MPa at 35 °C, 10 min and a 67% CO2 volume ratio) and to conventional thermal treatment (95 °C/1 min). Physicochemical properties (pH, soluble solid content, ζ-potential, particle size distribution and rheological behavior), organic acid (citric and malic) content, and phenolic compounds (UHPLC-ESI-MS/MS) were evaluated; moreover, antioxidant activity assays (DPPH and TEAC) as well as sugar content and fructo-oligosaccharide analyses (HPAEC-PAD) were performed. The increase in pressure levels reduced the particle size suspended in the functional juice. Supercritical processing was able to preserve all compounds responsible for the functional properties of the beverage as well as the natural nutritional components such as sorbitol, glucose, fructose and sucrose. The emerging technology did not reduce the antioxidant activity of the juice samples, thus maintaining their functionality. The inulin chemical profile was not altered by the supercritical CO2 processing, while in the thermally treated sample, there was a breakdown of the inulin chain into units of short-chain fructo-oligosaccharides. Overall, supercritical technology may be an interesting option for inulin-enriched apple juice processing.

Biorefinery of turmeric (Curcuma longa L.) using non-thermal and clean emerging technologies: an update on the curcumin recovery step.

In this study, a biorefinery for the processing of turmeric (Curcuma longa L.) based on clean and emerging technologies has been proposed. High-intensity ultrasound (HIUS) technology was evaluated as a promising technique for curcumin recovery aiming to improve its extraction yield and technological properties as a colorant. In addition, we evaluated the effects of process conditions on the turmeric biomass after the extractions. The process variables were the number of stages of extraction with ethanol (1, 3 and 5) and the solvent to feed ratio (S/F) of 3, 5, 7, 9 (w/w). The highest curcumin content (41.6 g/100 g extract) was obtained using 1 wash and a S/F of 5 w/w, while the highest curcumin yield (3.9 g/100 g unflavored turmeric) was obtained using 5 stages and a S/F of 7. The extracts obtained by solid-liquid extraction assisted by HIUS showed a yellow color (157 and 169 of yellowness index) more intense than those obtained by the pressurized liquid extraction technique (101 of yellowness index) and better yield results than low-pressure solid-liquid extraction (using the same processing time). Thus, it was possible to obtain a characteristic yellow colorant with high curcumin yield in a short process time (5 min of extraction) using HIUS technology. Besides that, SEM images and FTIR spectra demonstrated that the turmeric biomasses processed by HIUS technology were not degraded.

Obtaining a novel mucilage from mutamba seeds exploring different high-intensity ultrasound process conditions.

We evaluated the effect of ultrasonic power (200-600 W) and process time (1-7 min) on the recovery of a novel polysaccharide from mutamba (Guazuma ulmifolia Lam.) seeds applying high-intensity ultrasound. Ultrasound process conditions intensification gradually was removing the mucilage layer around the hydrated seeds. Then, the scanning electron micrographs showed that the mucilage was removed completely at the highest applied energy density (10,080 J/mL). Although the colour of mutamba seed mucilage (MSM) have been changed due to increase of energy density, it not affects its practical use because the MSM can be purified to remove impurities. The results obtained in this study demonstrated that the ultrasound process conditions intensification did not affect the primary structure of MSM according to ζ-potential, FTIR spectrum, and monosaccharide residues data. In conclusion, ultrasound process conditions intensification allows the full recovery of the MSM at a short process time (7 min) without altering its quality and the primary structure.