Abstract The goal of this study was to evaluate the α-amylases production by basidiomycetes in submerged cultivation (SmC), using wheat milling by-products as substrate. Enzymatic activity was evaluated to select the best strain, the ideal concentration, the by-product and the influence of supplementation in the culture medium. The best producer was chosen among three strains, Ganoderma applanatum (MR-56), Schizophyllum commune (MR-01), and Ganoderma stipitatum (MR-72). All of them were cultivated under the same conditions. MR-56 was selected for an initial screening activity of 5.10±0.31 U/mL in 192h. The Ganoderma applanatum strain was cultured in a medium with various starch concentrations to determine which concentration resulted in the highest α-amylase activity. After 120h of cultivation, activity in the medium containing 4.5% (w/v) had increased from 0.40±0.03 U/mL to 20.70±0.9 U/mL. Subsequently, the wheat milling by-products [Glue Flour (GF), Clean-Out Flour (CF4) Flour, and Low-grade Flour (LGF)] were evaluated in aqueous suspension with and without nutritional supplementation. It was shown that GF and LGF are potential starch sources which do not require nutritional supplementation, exhibiting increases in α-amylase activity of 31.79 U/mL and 30.98 U/mL, respectively. This is the first report involving the application of Ganoderma applanatum in SmC for α-amylase production using synthetic and wheat milling by-products. The results also demonstrated that wheat milling by-products may also be used without supplementation for cell growth and enzyme production, contributing to the development of more sustainable forms of enzyme production through the innovative use of edible organisms with medicinal properties.

Abstract Background There are limited real-world data on the clinical course of untreated coronary lesions according to their functional severity. Objective To evaluate the 5-year clinical outcomes of patients with revascularized lesions with fractional flow reserve (FFR) ≤ 0.8 and patients with non-revascularized lesions with FFR > 0.8. Methods The FFR assessment was performed in 218 patients followed for up to 5 years. Participants were classified based on FFR into ischemia group (≤ 0.8, intervention group, n = 55), low-normal FFR group (> 0.8-0.9, n = 91), and high-normal FFR group (> 0.9, n = 72). The primary endpoint was major adverse cardiac events (MACEs), a composite of death, myocardial infarction, and need for repeat revascularization. The significance level was set at 0.05; therefore, results with a p-value < 0.05 were considered statistically significant. Results Most patients were male (62.8%) with a mean age of 64.1 years. Diabetes was present in 27%. On coronary angiography, the severity of stenosis was 62% in the ischemia group, 56.4% in the low-normal FFR group, and 54.3% in the high-normal FFR group (p<0.05). Mean follow-up was 3.5 years. The incidence of MACEs was 25.5%, 13.2%, and 11.1%, respectively (p=0.037). MACE incidence did not differ significantly between the low-normal and high-normal FFR groups. Conclusion Patients with FFR indicative of ischemia had poorer outcomes than those in non-ischemia groups. There was no difference in the incidence of events between the low-normal and high-normal FFR groups. Long-term studies with a large sample size are needed to better assess cardiovascular outcomes in patients with moderate coronary stenosis with FFR values between 0.8 and 1.0.

Resumo Fundamento Existem dados limitados sobre a evolução clínica de lesões coronarianas não tratadas de acordo com sua gravidade funcional no mundo real. Objetivo Este estudo teve como objetivo avaliar os resultados clínicos de até 5 anos em pacientes com lesões revascularizadas com reserva de fluxo fracionada (FFR) ≤ 0,8 e em pacientes com lesões não revascularizadas com FFR > 0,8. Métodos A avaliação pelo FFR foi realizada em 218 pacientes seguidos por até 5 anos. Os participantes foram classificados com base na FFR no grupo isquêmico (≤ 0,8, grupo intervenção, n = 55), no grupo FFR normal-baixa (> 0,8-0,9, n = 91) e no grupo FFR normal-alta (> 0,9, n = 72). O desfecho primário foram eventos cardíacos adversos maiores (ECAMs), um composto de morte, infarto do miocárdio e necessidade de nova revascularização. O nível de significância adotado neste estudo foi alfa = 0,05; deste modo, resultados com valores de p < 0,05 foram considerados estatisticamente significativos. Resultados A maioria dos participantes era do sexo masculino (62,8%) com média de idade de 64,1 anos. Diabetes estava presente em 27%. À angiografia coronariana, a gravidade da estenose avaliada foi de 62%, 56,4% e 54,3% nos grupos isquêmico, FFR normal-baixa e FFR normal-alta, respectivamente (p < 0,05). O período médio de acompanhamento foi de 3,5 anos. A incidência ECAM foi de 25,5%, 13,2% e 11,1%, respectivamente (p = 0,037). Não houve diferença na incidência de ECAM entre os grupos FFR normal-baixa e FFR normal-alta (p = NS). Conclusão Pacientes com FFR indicativa de isquemia apresentaram piores desfechos quando comparados aos dos grupos não isquêmicos. Entre os grupos que apresentaram valores de FFR considerados normal-baixo e normal-alto, não houve diferença na incidência de eventos. Há necessidade de estudos de longo prazo e com grande número de pacientes para melhor avaliar os desfechos cardiovasculares em pacientes portadores de estenose coronariana moderada com valores de FFR entre 0,8 e 1,0.

Resumen Los compuestos aromáticos tienen amplias aplicaciones en las industrias alimentaria, cosmética y farmacéutica. Sin embargo, estos compuestos se producen principalmente por síntesis química. Una alternativa, sostenible y prometedora para la producción natural de compuestos aromáticos son los procesos fermentativos usando microrganismos. En este trabajo se obtuvo la producción de diferentes compuestos aromáticos en concentraciones significativas, como acetato de isoamilo, alcohol isoamílico, acetato de etilo y acetaldehído, entre otros, a partir de subproductos de la agroindustria (melaza de caña de azúcar), usando Pichia fermentans como inoculo. Los compuestos aromáticos se concentraron posteriormente por pervaporación y microencapsulación mediante secado por aspersión utilizando diferentes agentes encapsulantes. Se encontró que cuando estas técnicas se aplicaron a dos bebidas, produjeron un olor de notas afrutadas, especialmente las de plátano. Además, los panelistas identificaron notas amaderadas y alcohólicas. Finalmente, se encontró que los mejores agentes encapsulantes eran la dextrina (73,49 ppm) y la mezcla de dextrina y maltodextrina (72,58 ppm).

Abstract Aroma and fragrance compounds have broad applications in the food, cosmetic and pharmaceutical industries. However, these compounds are mainly produced by chemical synthesis. An alternative, sustainable and promising route for the natural production of flavour compounds is microbial transformation processes. In this work, we obtained the production of different aroma compounds in significant concentrations, such as isoamyl acetate, isoamyl alcohol, ethyl acetate and acetaldehyde, among others, from agribusiness by-products (sugarcane molasses) and using Pichia fermentans. These were subsequently concentrated by pervaporation and microencapsulation by spray drying using different encapsulating agents. The results showed that when these techniques were applied to two iced beverages, they produced an odour of fruity notes, especially those of banana. In addition, woody and alcoholic notes were identified by the panellists. Finally, the best encapsulating agents were dextrin (73.49 ppm) and the dextrin and maltodextrin mixture (72.58 ppm).

Abstract The aim of this study was to evaluate the performance of Caspian Sea-style spontaneous milk fermentation to improve the quality of pasteurized milk containing high levels of Pseudomonas contamination, with a focus on microbiological safety and stability of the final product. Bacterial diversity of pasteurized milk, fermentation process, and after 60 days of storage was analyzed by Illumina-based sequencing, and presence of viable taxa was confirmed by culturing on selective media. Low quality pasteurized milk harbored mainly Gram-negative bacteria, markedly dominated by Pseudomonas. Following fermentation, lactic acid bacteria rapidly became dominant with maximum population of 10.15 log CFU/mL at 18 h, represented mainly by Lactococcus. However, sequences related to Pseudomonas, and to a lesser extent for enterobacteria, remained constant throughout the fermentation process. The cultured-dependent approach confirmed the presence of viable Pseudomonas, with a final population of 5.60 log CFU/mL. Biochemical transformations were further analyzed, indicating lactic acid as the main end-metabolite produced (maximum concentration of 5.93 g/L at 24 h). In addition, the increase of 2-nonanone can be correlated as a volatile biomarker of P. aeruginosa and related species. Altogether, the results demonstrated that natural milk fermentation may often not inhibit the development of pathogens and food spoilage microorganisms.

Abstract Food supplements have been increasingly investigated. Probiotics have several benefits for human and animal health and selenium (Se) is widely recommended against oxidative stress. In this context, the aim of this study was to develop a low-cost bioprocess to produce a functional food product comprising both probiotic and Se accumulation. Yeast cells of Saccharomyces boulardii CCT 4308 were cultivated using sugarcane molasses as substrate. Optimization studies were performed to evaluate the best medium composition for biomass production and Se-accumulation in batch and fed-batch systems. Optimized conditions were defined with a medium composed of 150 g L-1 sugarcane molasses and 12 g L-1 yeast extract, with feeding of 100 g L-1 sugarcane molasses and 100 μg mL-1 of Se incorporation after 4 h and 10 h of fermentation, respectively, during 48 h in STR (stirred tank reactor). Best biomass production reached 14.52 g L-1 with 3.20 mg Se g-1 biomass at 12 h. Process optimization led to 4.82-fold increase in biomass production compared to initial condition. A final Se-enriched S. boulardii CCT 4308 biomass was obtained, which is comparable to commercial products. An alternative probiotic yeast biomass was efficiently produced as a new food-form of Se supplement in a sustainable process using an inexpensive agro-industrial residue.

Abstract With the COVID-19 pandemic, many diagnostic tests (molecular or immunological) were rapidly standardised, given the urgency of the situation, many are still in the process of being validated. The main objective of this study was to review the aspects of the diagnostic kits approved in Brazil and their application in the different federative units to gather epidemiological information. In order to achieve these objectives, a survey was carried out on the data available at the regulatory agency (ANVISA) and in the literature. The main countries that have registered products in Brazil are China (51.4%), Brazil (16.6%), South Korea (9.2%), USA (8.8%) and Germany (3.6%). The methodologies of these products are based on the detection of nucleic-acid (15.8%), antigen (13%) and antibody (71.2%). In the immunological tests, it was verified that the sensitivity ranged from 55 to 100% and the specificity from 80 to 100%. The percentage of cases in the samples tested in Brazil is elevated in almost all federative units since eight states showed 40% of positive cases in tested samples, while 18 states displayed between 20 and 40%. In conclusion, this review showed that Brazil is dependent on external technology to respond to pandemics, epidemics and endemics disease and needs to improve its biotechnological scheme to solve further diseases outbreaks.

RESUMEN El objetivo de este estudio fue evaluar varios métodos de ruptura de las esporas de Ganoderma lucidum y extraer sus propiedades bioactivas. Para este propósito se evaluaron diferentes técnicas de rompimiento como: la maceración manual (RM), la maceración con esferas de diversos materiales (BR) y la exposición a microondas junto la maceración de las esporas con esferas de acero/cromo (MBR1). La ruptura de las esporas fue evaluada por espectroscopia UV-Vis, la cual mostró que las vibraciones 2955, 1642, 1240, 1080 y 1746 cm-1 correspondieron a cambios estructurales en las paredes de las esporas. El extracto MBR1 presento el mayor contenido de carbohidratos (19,80 mg.g-1) y polifenoles (2,21 mg.g-1), mientras que el extracto BR tuvo una mayor actividad antioxidante (57,22% Inb DPPH). Los extractos MBR1 y BR también presentaron en el análisis de monosacáridos un 62,2 y 73,5% de contenido glucosa. Como conclusión la mejor metodología para extraer biomoléculas de las paredes de las esporas de G. lucidum fueron el tratamiento térmico con microondas y la ruptura de las paredes con esferas de acero/cromo, porque este proceso permitió la extracción de una mayor cantidad de carbohidratos con posibles propiedades antioxidantes.

SUMMARY This study aimed at evaluating effective methods for breaking the hard and insoluble spores of Ganoderma lucidum to recover functional biomolecules. Rupture techniques were evaluated such as manual maceration (RM), maceration with spheres of various materials (BR), and microwave exposure plus maceration with steel/ chrome spheres (MBR1). Spore rupture was evaluated using UV-Vis spectroscopy, which showed vibrations of 2955, 1642, 1240, 1080 and 1746 cm-1 corresponding to changes in spore walls. The MBR1 extract contained the largest amounts of carbohydrates (19.80 mg.g-1 spores) and polyphenols (2.21 mg.g-1 spores), whereas the BR extract had higher antioxidant activity (57.22%Inb DPPH). The MBR1 and BR extracts contained 62.2 and 73.5% glucose, respectively. Both methods also involved significant extraction of carbohydrates and proteins. The best way to extract biomolecules from spore walls is to perform a microwave heat treatment and break the walls with steel/chrome spheres; this produces large quantities of carbohydrates with antioxidant properties.

Abstract We present an improved method of direct transesterification suitable for the quantitative analysis of multiple dry samples for its fatty acid content, using a minimal amount of biomass and reactants. The method features an acid-catalyzed direct alcoholysis of microgram samples of dry biomass; the rationale behind the solvent and reagent proportions chosen is discussed. The method was validated using seven microbial strains with diverse lipid content (Saccharomyces cerevisiae, Saccharomyces boulardii, Candida tropicalis, Haematococcus pluvialis, Chlorella vulgaris, Spirulina platensis and Schizochytrium limacinum), and compared with a macroscale direct transesterification method, and with gravimetric analysis of lipids extracted with solvents. The microscale method showed a conversion of 98.06 ± 0.87% of the lipids, using approximately 3 mg of dry biomass, 1mL of 0.2M H2SO4 dissolved in anhydrous ethanol (the acid is the catalyzer and ethanol the reactant)). The mixture was maintained at 70 °C for 20 h with periodic mixing, and then extracted with 2mL n-heptane and analyzed by GC-FID. The lipid content was then calculated considering dilution and sample mass. This method is effective, reliable, and technically attractive for analytical and comparative purposes.

ABSTRACT Nidularium procerum and Nidularium innocentii (Bromeliaceae) were cultivated in vitro on media supplemented with different sources and levels of GA3 (gibberellic acid). These sources were the commercial powder (analytical degree) and fermented extract obtained by Fusarium moniliforme via solid state fermentation. The in vitro elongation and rooting of these plants were evaluated after 50 days of cultivation. The GA3 present in the fermented extract possess the same effect of purified GA3 (analytical degree) for the increase of the height of aerial part of shoots of N. innocentii, but not for the N. procerum being the GA3 fermented extract in a lesser degree. The GA3 fermented extract influences negatively the rooting in N. innocentii, while GA3 analytical degree practically does not interfere in the rooting. On the other hand, in N. procerum, both the GA3 sources reduce the root number and do not interfere in rooting percentage. GA3 crude fermented extract is an alternative to reduce costs, however, its results can vary depending on the species and parameter evaluated. The fermented extract was stored at temperature during 260 days and its shelf life presented a suitable stability, maintaining 92% of its initial GA3 amount.

Abstract INTRODUCTION: The production of the Montenegro antigen for skin test poses difficulties regarding quality control. Here, we propose that certain animal models reproducing a similar immune response to humans may be used in the quality control of Montenegro antigen production. METHODS: Fifteen Cavia porcellus (guinea pigs) were immunized with Leishmania amazonensis or Leishmania braziliensis , and, after 30 days, they were skin tested with standard Montenegro antigen. To validate C. porcellus as an animal model for skin tests, eighteen Mesocricetus auratus (hamsters) were infected with L. amazonensis or L. braziliensis , and, after 45 days, they were skin tested with standard Montenegro antigen. RESULTS: Cavia porcellus immunized with L. amazonensis or L. braziliensis , and hamsters infected with the same species presented induration reactions when skin tested with standard Montenegro antigen 48-72h after the test. CONCLUSIONS: The comparison between immunization methods and immune response from the two animal species validated C. porcellus as a good model for Montenegro skin test, and the model showed strong potential as an in vivo model in the quality control of the production of Montenegro antigen.

L-Lysine is an essential aminoacid added as supplement for animal feed. The aim of this work was to produce an L-Lysine enriched bran using Brazilian agroindustrial byproducts. Both the raw material costs and purification steps were minimized. Firstly, medium composition for the growth of Corynebacterium glutamicum ATCC 21799 was optimized targeting enhanced L-Lysine production - salt, vitamins and nitrogen sources concentrations were tested and selected. Next, UV mutant strains were generated and the best producers were used in formulated media using sugarcane molasses. It was reached a production of 9.3 g/L of L-Lysine with the optimized formulated media. This L-Lyisne rich broth was then impregnated and cyclically reimpregnated in pre-treated solid matrixes (sugarcane bagasse, citrus pulp, brewer spent grain, soybean husk and wheat bran). After processing, it was generated enriched brans with significant amounts of L-Lysine (13.8%, 7.0%, 8.9%, 5.9% and 8.4%, respectively), which has an interesting market potential for animal feed.

The production of value-added products could be a valuable option for cheese wastewater management. However, this kind of study cannot just focus alone on getting the final product. This also necessitates studies on the dynamics of bioprocesses. With these as background, the present investigation aimed at evaluating the buffering capacity of deproteinized whey and effect of temperature and nitrogen source on ethanol yields from it. The batch fermentation conditions used to evaluate ethanol production were temperatures 30, 35, 40°C and pH 4.5, 5.0, 5.5, 6.0. To study the influence of nitrogen source on ethanol yield, a design matrix was applied using yeast extract and (NH4)2SO4.The final pH was analyzed to evaluate the buffering capacity. The results showed that the Kluyveromyces marxianus was thermotolerance to produce ethanol at 35 and 40°C, which was not observed at 30°C. Results also showed that the deproteinization procedure did not affect the buffering capacity of cheese whey. Finally, higher ethanol production was obtained using yeast extract (3% v/v). These results could be important for developing low-cost method for industrial production of ethanol from deproteinized whey.

This aim of study was to compare the performance of a biofilter (BF) and trickle bed reactor (TBR) under increased styrene loading with a constant acetone load, 2 gc/m3/h. At styrene loading rates up to 30 gc/m3/h, the BF showed higher styrene removal than TBR. However, the BF efficiency started to drop beyond this threshold loading and could never reach steady state, whereas the TBR continued to yield a 50% styrene removal. The acetone removal remained constant (93-98%) in both the reactors at any styrene loading. Once the overloading was lifted, the BF recovered within 26 min, whereas the TBR efficiency bounced back only to 95%, gradually returning to complete removal only in 10 h.

The aim of this study was to evaluate the probiotic properties of Pediococcus acidilactici B14 and to study its resistance in the gastrointestinal system when combined with Lactobacillus acidophilus ATCC 4356 and used in a potentially symbiotic aerated soy based dessert. P. acidilactici B14 showed some important probiotic characteristics such as survival rate of 45.9% at pH 2.5; 72.4% in 0.3% bile salts and 95.8% after gastrointestinal transit at pH 4.0. Tolerance against the antibiotics cephalexin, neomycin, vancomycin, cefotaxime and penicillin G was also observed. The strain inhibited antagonism against the following cultures: Escherichia coli ATCC 25922, Bacillus cereus ATCC 33018, Staphylococcus aureus ATCC 6538P and Salmonella sp. The mixed culture of P. acidilactici B14 with L. acidophilus ATCC 4356 showed a survival rate of 92.4% after the passage through the gastrointestinal system at pH 4.0. Furthermore, in the presence of the food matrix, an average increase in cell viability, after being subjected to the gastrointestinal system of 9.9% at pH 2.0 and 6.1% at pH 4.0, was observed. This characterized the adequacy of the associated culture as probiotic in the development of a functional food such as soy based aerated symbiotic dessert.

Poly-ß-hydroxybutyrate (PHB) is a biodegradable intracellular microbial product produced by many bacteria and it is comparable to some of the petrochemical derived thermoplastics such as polypropylene. One of the main barriers for the commercial exploitation is the high cost of the substrate for the production of biopolymer. The utilization of mixed microbial cultures facilitates the use of complex substrates thereby reducing the cost of PHB production. In the present study, mixed culture systems were evaluated for PHB production. Bacillus firmus NII 0830 was used for the production of PHB since it accumulates a large amount of PHB and a second organism Lactobacillus delbrueckii NII 0925 was used to provide lactic acid. FTIR and 1H NMR analyses revealed that the PHB extracted from pure culture and mixed culture showed exact match to that of standard PHB. Biodegradation studies of the PHB blends showed 87% degradation. It was also found that a consortium of organisms degraded the films faster than a single organism.