Pilot scale production of high-content mycoprotein using Rhizopus microsporus var. oligosporus by submerged fermentation and agro-industrial by-products.

While mycoprotein has gained traction as a human food source, its potential as a nutrient for animals remains largely unexplored. The mycoprotein-producing Rhizopus microsporus var. oligosporus, a fungus traditionally used for human food in Indonesia, is promising. It could revolutionise animal nutrition once it is Generally Recognized as Safe (GRAS) and is a biosafety level 1 (BSL1) organism. To enhance sustainably, we propose using sugar cane molasses (SM) and corn steep liquor (CSL) as nutrient sources. Also, we investigated the growth of R. microsporus var. oligosporus in five 14 L external-loop airlift bioreactors using CSL as the sole nutrient source. After 96 h of fermentation, at 25 °C and 0.5 vvm, the mycelium produced had an average biomass yield of 38.34 g L-1, with 70.18 % (m v-1) crude protein (mycoprotein). This bioprocess, which is scalable and economically viable, produces high amounts of mycoprotein for animal feed using CSL, a cost-effective agro-industrial by-product, providing a practical solution to the growing demand for animal protein.

Fungal Laccases and Fumonisin Decontamination in Co-Products of Bioethanol from Maize.

Maize (Zea mays L.) may be infected by Fusarium verticillioides and F. proliferatum, and consequently contaminated with fumonisins (FBs), as well as the co-products of bioethanol intended for animal feed. Laccase enzymes have a wide industrial application such as mycotoxin degradation. The aims were to isolate and identify fungal laccase-producing strains, to evaluate laccase production, to determine the enzymatic stability under fermentation conditions, and to analyse the effectiveness in vitro of enzymatic extracts (EEs) containing laccases in degrading FB1. Strains belonging to Funalia trogii, Phellinus tuberculosus, Pleurotus ostreatus, Pycnoporus sanguineus and Trametes gallica species showed laccase activity. Different isoforms of laccases were detected depending on the evaluated species. For the FB1 decontamination assays, four enzymatic activities (5, 10, 15 and 20 U/mL) were tested, in the absence and presence of vanillic acid (VA) and 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) as redox mediators (1 and 10 mM). Trametes gallica B4-IMICO-RC EE was the most effective strain in buffer, achieving a 60% of FB1 reduction. Laccases included in EEs remained stable at different alcoholic degrees in maize steep liquor (MSL), but no significant FB1 reduction was observed under the conditions evaluated using MSL. This study demonstrate that although laccases could be good candidates for the development of a strategy to reduce FB1, further studies are necessary to optimise this process in MSL.

The in vitro rumen exsheathment test for studying the effect of plant extracts on the exsheathment of Haemonchus contortus infective larvae.

This study applied the in vitro rumen exsheathment test (IVRET) to evaluate the exsheathment kinetics of Haemonchus contortus infective larvae (L3) incubated in ruminal liquor (RL) containing acetone:water extracts of Acacia pennatula (AP), Gymnopodium floribundum (GF), Havardia albicans (HA) or Lysiloma latisiliquum (LL). The role of polyphenols in the biological activity of the evaluated extracts was also determined. Larvae were incubated in RL either alone or added with a different plant extract (AP, GF, HA, or LL) at 1200 µg/mL. Polyethylene glycol (PEG) was added to block polyphenols in each treatment (RL+PEG, AP+PEG, GF+PEG, HA+PEG, and LL+PEG). After incubation times of 0, 1, 3, 6, 9, and 24 h, the exsheathment process was stopped to count the number of ensheathed and exsheathed L3. A Log-Logistic model was used to determine the L3 exsheathment kinetics in the different RL treatments. The inflection point of the respective kinetic curves, which indicates the time to reach 50 % exsheathed L3 (T50), was the only parameter that differed when comparing the exsheathment models (99 % probability of difference). The T50 values obtained for GF, HA, and LL treatments (T50 = 7.11 - 7.58 h) were higher in comparison to the T50 of RL (5.72 h) (≥ 70 % probability of difference). The L3 incubated in RL added with GF, HA, and LL extracts delayed their exsheathment at 3 and 6 h of incubation (28.71 - 48.06 % exsheathment reduction) compared to the RL treatment. The T50 value for AP, AP+PEG, GF+PEG, HA+PEG, and LL+PEG were similar to RL and RL+PEG (T50 = 5.34 - 6.97 h). In conclusion, the IVRET can be used to identify plants with the potential to delay the exsheathment of H. contortus L3 in the ruminal liquor. The acetone:water extracts of G. floribundum, H. albicans, and L. latisiliquum delayed the T50 of H. contortus exsheathment, which was evident at 3 and 6 h of incubation in ruminal liquor. The observed exsheathment delay was attributed to the polyphenol content of the extracts.

Adapting the in vitro rumen incubation method to evaluate the effect of a plant extract on the exsheathment inhibition of Haemonchus contortus infective larvae.

This study adapted the in vitro rumen incubation (IVRI) method to evaluate the biological activity of a Gymnopodium floribundum leaves extract against the exsheathment of Haemonchus contortus infective larvae (L3), and to determine the role of plant polyphenols on the biological activity. The incubation protocol followed the IVRI method, adding polyethylene glycol (PEG) as a polyphenol-blocking agent. The L3 were incubated in ruminal liquor (RL), ruminal liquor with PEG (RL+PEG), ruminal liquor with G. floribundum extract (RLE), and ruminal liquor with G. floribundum extract and PEG (RLE+PEG). Incubation condition controls included phosphate buffered saline (PBS), PBS with PEG (PBS+PEG), incubation medium (without ruminal liquor) (IM), and incubation medium with PEG (IM+PEG). The L3 were recovered after incubation times of 0, 1, 3, 6, 9, and 24 h (39 °C). The respective L3 exsheathment kinetics were estimated for the different treatments (RL, RL+PEG, RLE, and RLE+PEG) using Log-Logistic models. The parameters of the different models were compared to determine the impact of the extract, with or without PEG, on the L3 exsheathment kinetics. The exsheathment in PBS and PBS+PEG remained < 2.71% at each incubation time. The exsheathment in IM and IM+PEG reached 13.58% and 17.18% at 24 h, respectively. The exsheathment percentages for RLE were lower than those for RL at 3, 6 and 9 h of incubation. The inflection point, indicating the time required to reach 50% of the maximal exsheathment (T50), was the only parameter that differed between the ruminal liquor models. The T50 in RLE (7.106 h) was higher than the values obtained for RL (5.385 h) and RL+PEG (4.923 h) (99.99% probability of being different). Such delay resulted in a reduction of exsheathment in RLE of 62% at 3 h, 38% at 6 h, and 12% at 9 h, relative to RL values. When PEG was added with the extract (RLE+PEG), the T50 (5.045 h) was similar to that of RL and RL+PEG. The IVRI method was adapted as an in vitro rumen exsheathment test (IVRET). The IVRET showed that H. contortus L3 exposed to G. floribundum extract delayed their exsheathment kinetics at different time points. The exsheathment delay was attributed to the polyphenol content of the extract.

Raman Spectroscopic and Sensory Evaluation of Cocoa Liquor Prepared with Ecuadorian Cocoa Beans Treated with Gamma Irradiation or Induced Electromagnetic Field Fermentation.

Cocoa liquor is the primary precursor of the worldwide highly appreciated commodity chocolate. Its quality depends on several factors, such as the type of cocoa, the fermentation process, and the control of the contaminants in the fermented beans. This study aims to evaluate whether the induced magnetic field treatment during the fermentation process or the pathogen reduction with gamma irradiation after the fermentation affect the characteristics of the cocoa liquor obtained from Ecuadorian cocoa beans. For this purpose, liquor samples from controls (standard process), from beans treated with an induced magnetic field up to 80 mT, and from beans irradiated with nominal doses up to 3 kGy were characterized through Raman spectroscopic analysis and sensorial evaluation. The most relevant bands of the cocoa liquor were assigned according to reports from the literature, spectroscopic data, and chemometrics. The spectra corresponding to different treatments and doses were visually very similar, but they could be discriminated using OPLS-DA models, where the most intense Raman signals were attributed to the lipid components. The sensorial evaluation rated the presence of floral, fruity, almondy, acid, and bitter flavors, along with astringency and intense aroma, and these attributes exhibited variable behavior depending on the dose of the irradiation or magnetic treatment. Therefore, both treatments may exert an influence on cocoa beans and, therefore, on the cocoa liquor quality.

Optimized fed-batch cultivation of Rhodotorula toruloides in a bubble column bioreactor progressed the ß-carotene production from corn steep liquor.

In this work, a new isolate yeast, namely Rhodotorula toruloides KP324973, was examined for ß-carotene production from corn steep liquor (CSL) as a sole carbon source because CSL as the by-product of corn wet-milling process mainly enriched from the water-soluble carbohydrates. The studies were preliminary performed at the shaken flasks, and then developed at batch and fed-batch modes in a bubble column reactor (BCR). Application of the BCR improved the carotenogenesis of the cells in comparison with shaken flasks and the specific ß-carotene production rate (Rp) and the yield of ß-carotene production from the total reducing sugars (YP/TRS) reached 2.23 mg gcell-1 h-1 and 36.82 mg gTRS-1, respectively. Further studies were carried out to optimize the operational factors of the BCR for a fed-batch production by the response surface methodology. An optimal condition at a feed flow rate of 2.5 mL h-1, temperature 11.7°C, and initial pH of 6.1 obtained the highest Rp = 12.31 mg gcell-1 h-1 and YP/TRS = 97.18 mg gTRS-1.

Improvement of DOPA-Melanin Production by Aspergillus nidulans Using Eco-Friendly and Inexpensive Substrates.

Fungal pigments, including melanin, are recognized as promising materials for biomedical, environmental, and technological applications. In previous studies, we have demonstrated that the DOPA-melanin produced by the MEL1 mutant of Aspergillus nidulans exhibits antioxidant, anti-inflammatory, and antimicrobial activities without any cytotoxic or mutagenic effects, suggesting its potential use in pharmaceuticals. In order to increase the yield of this pigment and reduce the costs of its large-scale production, the present study aimed to evaluate agro-industrial by-products, sugarcane molasses, vinasse, and corn steep liquor as inexpensive substrates for fungal growth using experimental design methodology. According to the results obtained, the optimal composition of the culture medium was 0.81% (v/v) vinasse and 1.62% (w/v) glucose, which promoted a greater production of melanin (225.39 ± 4.52 mg g-1 of biomass), representing a 2.25-fold increase compared with the condition before optimization (100.32 mg.g-1 of biomass). Considering the amount of biomass obtained in the optimized condition, it was possible to obtain a total melanin production of 1 g L-1. Therefore, this formulation of a less complex and low-cost culture medium composition makes the large-scale process economically viable for future biotechnological applications of melanin produced by A. nidulans.

Lignin-Degrading Bacteria in Paper Mill Sludge.

The effluents generated in the paper industry, such as black liquor, have a high content of lignin and other toxic components; however, they represent a source of lignin-degrading bacteria with biotechnological potential. Therefore, the present study aimed to isolate and identify lignin-degrading bacteria species in paper mill sludge. A primary isolation was carried out from samples of sludge present in environments around a paper company located in the province of Ascope (Peru). Bacteria selection was made by the degradation of Lignin Kraft as the only carbon source in a solid medium. Finally, the laccase activity (Um-L-1) of each selected bacteria was determined by oxidation of 2,2'-azinobis-(3-etilbencenotiazolina-6-sulfonate) (ABTS). Bacterial species with laccase activity were identified by molecular biology techniques. Seven species of bacteria with laccase activity and the ability to degrade lignin were identified. The bacteria Agrobacterium tumefasciens (2), Klebsiella grimontii (1), and Beijeinckia fluminensis (1) were reported for first time. K. grimowntii and B. fluminensis presented the highest laccase activity, with values of 0.319 ± 0.005 UmL-1 and 0.329 ± 0.004 UmL-1, respectively. In conclusion, paper mill sludge may represent a source of lignin-degrading bacteria with laccase activity, and they could have potential biotechnological applications.

Caracterización física y sensorial de 24 genotipos especiales de cacao Theobroma cacao

Colombia, se destaca como país productor de cacao fino y de aroma, por lo cual, resulta de gran importancia explorar las características físicas y sensoriales de la diversidad genética existente. Para el presente estudio, se seleccionaron 22 genotipos de colecciones de trabajo con atributos productivos sobresalientes y dos testigos comerciales, reconocidos por su productividad y calidad sensorial; las muestras evaluadas fueron tomadas, de acuerdo con un muestreo aleatorio simple y se realizó un proceso de poscosecha homogéneo, controlado e independiente por genotipo, para obtener muestras de cacao seco individuales. Se realizaron análisis físicos al grano de cada genotipo y se preparó licor de cacao con cada una de las muestras obtenidas. Posteriormente, se evaluó cada licor por medio del panel de evaluación sensorial entrenado, donde se identificó y cuantificó la intensidad de atributos básicos, atributos especiales y atributos adquiridos, característicos de cada muestra. La información fue analizada mediante estadística descriptiva y análisis de componentes principales, lo que permitió la identificación de tres genotipos sobresalientes por el índice de grano y el porcentaje de cascarilla. El análisis sensorial evidenció que, en diecisiete genotipos, predominaron los atributos especiales, como herbal, floral, frutal, frutos secos y dulce. Estos resultados son un aporte importante para la selección de nuevas variedades de alta productividad, con características sensoriales de interés para la comercialización, que pueden ser evaluadas en diferentes regiones, para aumentar la disponibilidad genética en futuros programas de renovación y siembra de cacao que está en expansión, en los distintos territorios del país.

Colombia stands out as a country that produces fine and aroma cocoa; therefore, it is of great importance to explore the physical and sensory characteristics of the existing genetic diversity. For the present study, 22 genotypes from working collections with outstanding productive attributes and two recognized commercial controls were selected and evaluated for its productivity and sensory quality; the evaluated samples were taken according to a simple random sampling and a homogeneous, controlled, and independent post-harvest process was carried out by genotype, to obtain individual dry cocoa samples. Physical analyzes were carried out on the grain of each genotype and cocoa liquor was prepared with each of the samples obtained. Subsequently, each liquor was evaluated by means of the trained sensory evaluation panel, where the intensity of basic attributes, special attributes and acquired attributes characteristic of each sample was identified and quantified. The information was analyzed through descriptive statistics and principal component analysis, which allowed the identification of three outstanding genotypes by the grain index, and the husk percentage. The sensory analysis showed that, in seventeen genotypes, special attributes predominated such as: herbal, floral, fruity, dried fruit and sweet. These results are an important contribution for the selection of new high productivity varieties with sensory characteristics of interest for commercialization, which can be evaluated in different regions, to increase genetic availability in future renewal and planting programs of cocoa that is expanding in the different territories of the country.

Forastero and Criollo cocoa beans, differences on the profile of volatile and non-volatile compounds in the process from fermentation to liquor.

Cocoa bean fermentation is an important process because during this process, aroma compounds are produced, the astringency decreases, and the embryo dies. The fermentation processes of the Criollo and Forastero types have been studied separately without comparing them at the same time and in the same place. The aim of this work was to determine differences in the profile of volatile and nonvolatile compounds of Criollo and Forastero cocoa from the fermentation process to the final stage of obtaining the liquor. The experiments were carried out at the same time in the Maya region. Volatile compounds were determined by HS-SPME GC-MS (headspace solid phase-microextraction with gas chromatography-mass spectrometry). Sugars, organic acids, and alkaloids were determined by ultrahigh-performance liquid chromatography (UHPLC-PDA/UV). Criollo cocoa liquor was defined by the volatile and nonvolatile compounds such as acetic acid, phenylethyl alcohol, benzaldehyde, 2-phenylethyl acetate, acetophenone and 3-methylbutanal., which are associated with sour, honey, almond, flowery and chocolate aroma. Forastero cocoa liquor was represented with a significant difference by acetic acid, isobutyl acetate, 2,3-diethyl-5-methylpyrazine and ethyl octanoate and these could provide aroma descriptors such as sour, fruity and nutty. This study characterized for the first time the dynamics of volatile compounds during the fermentation, drying, and roasting stages and in the final cocoa liquor of Criollo and Forastero from cocoa beans of the same origin.

Nasal Cerebrospinal Leaks in the Milieu of COVID-19 Pandemic / Vazamentos nasais cerebroespinhais no meio da pandemia de COVID-19

Background The unintentional ingestion of oropharyngeal or gastric contents into the respiratory tract is known as aspiration. Rhinorrhea can cause aspiration pneumonia (cerebrospinal fluid leakage). Objective There are only a few reports in the literature about pneumonia as a complication of rhinorrhea. There are no reports on how to handle such cases if they present to the clinic at the peak of COVID-19 disease and distinguish between these two conditions. Methods We reviewed the literature and retrospectively analyzed the clinical information and treatment protocols used to treat the two clinical cases. Results By screening the COVID-19 PCR and antibodies more than twice, surgery was postponed for 10­14 days in both cases to rule out COVID-19-induced pneumonia. Chest CT scans still revealed ground glass opacities. In both cases, the skull base defect was repaired. In both cases, radiological signs of rhinorrhea-induced pneumonia had completely resolved at the 24- and 30-day follow-ups. Conclusion CSF aspiration causes radiological changes in the lungs in rhinnorhea. This is a short-term local decrease in lung tissue airness (partial filling of alveoli with fluids), which is visible radiographically (ground-glass opacities). To rule out COVID-19 infection, surgery should be postponed for 10­14 days, and PCR and antibodies (IgG, IgM) should be performed at least twice. If the COVID-19 screening test is negative, repair surgery can be scheduled.

Introdução A ingestão não intencional de conteúdo orofaríngeo ou gástrico no trato respiratório é conhecida como aspiração. A rinorréia pode causar pneumonia por aspiração (vazamento de líquido cefalorraquidiano). Objetivo Existem poucos relatos na literatura sobre pneumonia como complicação da rinorréia. Não há relatos sobre como lidar com esses casos se eles se apresentarem à clínica no pico da doença COVID-19 e distinguirem entre essas duas condições. Métodos Revisamos a literatura e analisamos retrospectivamente as informações clínicas e os protocolos de tratamento utilizados para tratar os dois casos clínicos. Resultados Ao rastrear a PCR e os anticorpos da COVID-19 mais de duas vezes, a cirurgia foi adiada por 10 a 14 dias em ambos os casos para descartar pneumonia induzida pela COVID-19. A tomografia computadorizada de tórax ainda revelou opacidades em vidro fosco. Em ambos os casos, o defeito na base do crânio foi reparado. Em ambos os casos, os sinais radiológicos de pneumonia induzida por rinorréia foram completamente resolvidos nos acompanhamentos de 24 e 30 dias. Conclusão A aspiração do LCR causa alterações radiológicas nos pulmões na rinorreia. Esta é uma diminuição local de curto prazo na leveza do tecido pulmonar (preenchimento parcial dos alvéolos com fluidos), que é visível radiograficamente (opacidades em vidro fosco). Para descartar infecção por COVID-19, a cirurgia deve ser adiada por 10 a 14 dias e a PCR e anticorpos (IgG, IgM) devem ser realizados pelo menos duas vezes. Se o teste de rastreio da COVID-19 for negativo, pode ser agendada uma cirurgia reparadora.

Immobilization and Application of the Recombinant Xylanase GH10 of Malbranchea pulchella in the Production of Xylooligosaccharides from Hydrothermal Liquor of the Eucalyptus (Eucalyptus grandis) Wood Chips.

Xylooligosaccharides (XOS) are widely used in the food industry as prebiotic components. XOS with high purity are required for practical prebiotic function and other biological benefits, such as antioxidant and inflammatory properties. In this work, we immobilized the recombinant endo-1,4-ß-xylanase of Malbranchea pulchella (MpXyn10) in various chemical supports and evaluated its potential to produce xylooligosaccharides (XOS) from hydrothermal liquor of eucalyptus wood chips. Values >90% of immobilization yields were achieved from amino-activated supports for 120 min. The highest recovery values were found on Purolite (142%) and MANAE-MpXyn10 (137%) derivatives, which maintained more than 90% residual activity for 24 h at 70 °C, while the free-MpXyn10 maintained only 11%. In addition, active MpXyn10 derivatives were stable in the range of pH 4.0−6.0 and the presence of the furfural and HMF compounds. MpXyn10 derivatives were tested to produce XOS from xylan of various sources. Maximum values were observed for birchwood xylan at 8.6 mg mL−1 and wheat arabinoxylan at 8.9 mg mL−1, using Purolite-MpXyn10. Its derivative was also successfully applied in the hydrolysis of soluble xylan present in hydrothermal liquor, with 0.9 mg mL−1 of XOS after 3 h at 50 °C. This derivative maintained more than 80% XOS yield after six cycles of the assay. The results obtained provide a basis for the application of immobilized MpXyn10 to produce XOS with high purity and other high-value-added products in the lignocellulosic biorefinery field.

Chocolates with Brazilian cocoa: Tracking volatile compounds according to consumers' preference.

This study aimed to evaluate the volatile compounds of chocolates made of Brazilian cocoas and statistically track them according to the products' sensorial profile in order to relate them to consumers' acceptance by preference map methodology. The intensity of the chocolate, acidity, woody, smoked, green, floral, burned, musty, and cocoa notes from chocolates produced with cocoa from different Brazilian states were analyzed by a trained panel and by 128 consumers. Samples from Côte d'Ivoire, which is known for its high-quality chocolate, were evaluated for comparison. Solid-phase microextraction headspace sampling/gas chromatography-mass spectrometry was employed to evaluate the samples' volatile compounds. One hundred volatile compounds were identified within the samples. The results from the preference maps showed that the maximum preference was found for chocolate made of cocoa from Rondônia, Bahia, and Espírito Santo and Côte d'Ivoire and organic samples from Pará. The ideal sample point was characterized by intense chocolate, floral, and woody notes and mild green and burned notes. The presence of furfural, 3-methyl butanal, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, methyl pyrazine, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, and tetramethyl pyrazine were shown to be important for consumer acceptance in the ideal product, whereas the presence of (Z)-2-heptenal and 2-pentyl furan may increase consumer rejection. 2,3-Methyl pyrazine, methyl pyrazine, and 2,3-butanediol, which are important volatile compounds previously reported in the literature, were statistically tracked to both positive and negative sample attributes and must be better explored concerning consumers' acceptance of chocolates.

Synthesis of a biobased resin and its screening as an alternative adsorbent for organic and inorganic micropollutant removal.

The sol-gel route was used to synthesize a biophenolic resin from a blend of Kraft black liquor and condensed tannin. The biobased resin has an amorphous structure and diversified surface functional groups. The biomaterial thermal stability was improved by Kraft black liquor, which increased the fixed carbon yield by 19.78% in an oxidant medium and 9.07% in an inert medium. Moreover, the presence of fixed carbon and char is positively related to the material flame retardant property. Additionally, impedance measurements were used to understand the physical phenomena occurring at the polymeric matrix's interface and the material's final properties. The biobased resin characterization and the considerable increase in the presence of micropollutants in surface and water bodies suggest the new biomaterial application in the adsorption process. Thus, its adsorption capacity toward several organic and inorganic micropollutants and its effectiveness in complex water matrices were evaluated. Methylene blue was used as a model compound to assess the influence of the resin composition on the adsorption capacity, and the type H isotherm indicates the high affinity of the biobased resin toward the micropollutant. The adsorption occurs in multilayer by intermolecular interaction and electrostatic forces. The amount of Kraft black liquor favored the adsorption, and the adsorption capacity was greater than 1250 mg g-1. When inorganic compounds were evaluated, the carboxyl and phenol groups favor the biomaterial affinity toward metal ions. Cu2+ and Ni2+ were completely removed from the contaminated water, and the adsorption capacity of the other inorganic compounds was: Pb2+ (36.97 mg g-1), Al3+ (22.17 mg g-1), Ba2+ (12.76 mg g-1), Ag1+ (33.85 mg g-1), and Fe2+ (19.44 mg g-1). In contrast, the adsorption capacity of the organic micropollutants was: 2,4-D (3.09 mg g-1), diuron (5.89 mg g-1), atrazine (2.71 mg g-1), diclofenac (2.04 mg g-1), caffeine (5.79 mg g-1), acetaminophen (4.80 mg g-1), methylene Blue (106.66 mg g-1), and methyl orange (30.48 mg g-1). The results pointed that the adsorption efficiency of organic micropollutants increases with the distribution coefficient (logD), indicating the biobased resin affinity toward more lipophilic compounds and ionized species.

Biosorption of nickel, cobalt, zinc and copper ions by Serratia marcescens strain 16 in mono and multimetallic systems.

The metallurgical industry is one of the main sources of heavy metal pollution, which represents a severe threat to life. Metals can be removed from aqueous solutions by using microbial biomasses. This paper analyses the heavy metal biosorption capacity of Serratia marcescens strain 16 in single and multimetallic systems. The results obtained show that Co(II), Ni(II) and Zn(II) biosorption in monometallic systems is two to three times higher than in the presence of bi-metallic and multimetallic solutions. Fourier transform infrared spectroscopy confirmed that carbonyl, carboxyl and hydroxyl were the main functional groups, as well as the amide bands I and II involved in metal uptake, which are present in external structures of the bacterial cell. The results obtained demonstrated the viability of S. marcescens strain 16 as a biosorbent for the design of eco-friendly technologies for the treatment of waste liquor.

Forward Black Liquor Acid Precipitation: Lignin Fractionation by Ultrafiltration.

Lignin recovery from black liquor is an important task for producing valuable chemical products. Acidification processes are currently applied by pulp and paper industries for black liquor treatment, in which two main streams are produced: the precipitated lignin fraction and a lignin-lean black liquor. Membrane filtration is a suitable alternative for lignin recovery from black liquor, but studies on lignin-lean black liquor filtration are scarce. Here, we evaluated the ultrafiltration process for lignin recovery from the both fractions of black liquor acidification. The lignin-lean black liquor presented 22 wt% of total solids with 4.6 wt% of lignin. Lignin retention from the lignin-lean black liquor by the 5 kDa ultrafiltration membrane was equal to 85%, with reduction in total solid concentration from 219.8 to 68.1 g L-1. Due to the relatively high solid concentration in the lignin-lean black liquor, cake formation was the main fouling mechanism during ultrafiltrations. The precipitated lignin solution presented 4.8 wt% of total solids with equivalent lignin concentration (4.7 wt%). The used membrane was able to retain almost 100% of solids and lignin from the solution prepared from the precipitated lignin. All fouling mechanisms were responsible for flux decay in ultrafiltration of the precipitated lignin solution. Steady state fluxes for lignin-lean black liquor and precipitated lignin solution were 0.9 and 15.9 L h-1 m-2, respectively. According to TGA analyses up to 800 °C, precipitated lignin and lignin-lean black liquor presented total mass losses of 63.5% and 44.3%, respectively. Also, the permeate samples presented lower mass losses than their respective feed samples. The ultrafiltration process reduced the average weight molar mass (Mw) of the precipitated lignin solution and lignin-lean black liquor from 1817 to 486 g mol-1and from 2876 to 1095 g mol-1, respectively. Thus, the 5 kDa ultrafiltration membrane was efficient for lignin recovery from the lignin-lean black liquor, while membranes with lower cut-off should be proposed for lignin purification from the precipitated fraction.

Sustainable Lipase Production by Diutina rugosa NRRL Y-95 Through a Combined Use of Agro-Industrial Residues as Feedstock.

The potential use of alternative culture media towards the development of a sustainable bioprocess to produce lipases by Diutina rugosa is clearly demonstrated. First, a synthetic medium containing glucose, peptone, yeast extract, oleic acid, and ammonium sulfate was proposed, with lipase activity of 143 U/L. Then, alternative culture media formulated with agro-industrial residues, such as molasses, corn steep liquor (CSL), and olive mill waste (OMW), were investigated. An experimental design was conducted, and only CSL concentration was found to have a positive effect in lipase production. The highest lipase activity (561 U/L) was produced on a mixture of molasses (5 g/L), CSL (6 g/L), OMW (0.5% v/v), 0.5 g/L of ammonium sulfate, and 3 g/L of peptone at 24 h of cultivation. Lipase production was also carried out in a 1-L bioreactor leading to a slightly higher lipase activity at 24 h of cultivation. The semi-purified enzyme exhibits an optimum temperature and pH of 40 °C and 7.0, respectively. Finally, the media cost per unit of lipase produced (UPC) was influenced by the medium components, specially by the inducer used. The lowest UPC was obtained when the agro-industrial residues were combined and used at the improved concentrations.

Hydrogen production by dark fermentation using a new low-cost culture medium composed of corn steep liquor and cassava processing water: Process optimization and scale-up.

The use of effluents for hydrogen production through dark fermentation is promising because it results in the generation of value-added products and reduction of the effluent's organic load. A low-cost medium using agroindustrial effluents, corn steep liquor (CSL) and cassava processing wastewater (CPW) was evaluated for hydrogen production with microbial consortia (Vir and Gal). Four variables were evaluated for their impact on biohydrogen production through a Plackett Burman design. Subsequently, the significant variables were optimized using a central composite design, resulting in two mathematical models with regression coefficients R2 > 0.92. The maximum yields were validated and resulted in 107 and 83.1 mL of biohydrogen/g COD removed for Vir and Gal, respectively. The lower medium cost for biohydrogen production was 81.5 USD/m3, approximately 80% more economical than some supplemented media. Finally, the scale-up of the biohydrogen production by consortia to 5L resulted in an increase of more than 40%.

Determinação espectrofotométrica de Cobre(II) em aguardente de mandioca (Tiquira) / Spectrophotometric determination of Copper (II) in manioc spirit (Tiquira)

RESUMO A pesquisa descreve a reação espectrofotométrica do complexo Cu(II)-PAN (1-(2-piridilazo)-2-naftol) e determina a concentração de cobre (II) em aguardente de mandioca (Tiquira). Para isso, dimensionamos a reação para os estudos de absorção do complexo, pH, estabilidade, proporção estequiométrica, interferentes, correlação e recuperação. Após essa etapa, determinamos a concentração de cobre(II) em cinco amostras de bebidas destiladas artesanalmente de cinco municípios do Estado do Maranhão. O resultados mostram que o complexo absorve em 548 nm em pH 6, é estável em um tempo de 7 horas em proporções estequiométrica mínima de 1:1 do ligante e metal, possui limite de detecção em 3,3 X 10-2 mg L-1, faixa de linearidade de 1 X 10-6 a 1 X 10-5 mol L-1, correlação (R2= 0,9990), desvio padrão=0,0035 e recuperação em 104,54 ± 7,01 e os principais íons interferentes são o ferro (III) e o cobalto (II). Na aguardente, constatamos que o teor de cobre está acima do permitido pela legislação brasileira.

SUMMARY The search describes the spectrophotometric reaction of the Cu (II)-PAN (1-(2-pyridilazo) -2-naphthol) complex and determines the concentration of copper (II) in manioc spirit (Tiquira). For this, we dimensioned the reaction for the studies of complex absorption, pH, stability, stoichiometric ratio, interferences, correlation, and recovery. After this step, we determined the concentration of copper (II) in five samples of beverages distilled artisanal from five municipalities of the State of Maranhão. The results show that the complex absorbs at 548 nm at pH 6, is stable at a time of 7 hours in minimum 1:1 stoichiometric proportions of the binder and metal, has a detection limit in 3.3 X 10-2 mg L-1, range (R2= 0.9990), standard deviation= 0.0035 and recovery at 104.54 ± 7.0, and the main interfering ions are iron (III) and cobalt (II). In the brandy, we find that the copper content is above that allowed by Brazilian legislation.

Cerebrospinal fluid analysis in 58 ruminants showing neurological disorders / Análise do líquido cefalorraquidiano em 58 ruminantes com distúrbios neurológicos

Ruminants may be affected by a wide variety of central nervous system (CNS) diseases. Cerebrospinal fluid (CSF) analysis forms the basis for ante mortem diagnostic evaluation of ruminants with clinical signs involving the CNS. Despite its importance as a tool to aid diagnosis, data regarding CSF examinations in spontaneous cases of CNS diseases in ruminants from Brazil are limited, and most reports involve experimental studies. Therefore, this study aimed to report the results of CSF analysis in 58 ruminants showing signs of neurological disorders. CSF samples for analysis were obtained from 32 cattle, 20 sheep, and 6 goats by cerebello-medullary cistern (n=54) or lumbosacral space (n=4) puncture. These ruminants showed neurological signs related to viral (n=13), mycotic (n=3), or bacterial (n=15) infections, and toxic (n=21), traumatic (n=4), or congenital disorders (n=2). CSF analysis from ruminants with viral infections presented lymphocytic pleocytosis, even though CSF showed no changes in several cases of rabies. Neutrophilic pleocytosis, cloudiness, presence of fibrin clots, and abnormal coloration were evident in the CSF of most cases of CNS bacterial infection, such as meningoencephalitis, meningitis, abscesses, myelitis, and a case of conidiobolomycosis. On the other hand, CSF was unchanged in most cases of toxic disorders, as botulism and hepatic encephalopathy. Elevated CSF density was observed in 60% of ruminants diagnosed with polioencephalomalacia. Our findings show that evaluation of CSF is a valuable diagnostic tool when used in association with epidemiological, clinical and pathological findings for diagnosis of CNS diseases in ruminants.(AU)

Os ruminantes podem ser afetados por uma grande variedade de doenças do sistema nervoso central (SNC). A análise do líquido cefalorraquidiano (LCR) constitui a base da avaliação diagnóstica ante mortem de ruminantes com sinais clínicos envolvendo o SNC. Apesar de sua importância como ferramenta para auxiliar no diagnóstico, os dados referentes aos exames do LCR em casos espontâneos de doenças do SNC em ruminantes no Brasil são limitados, e, a maioria dos relatos envolve estudos experimentais. Portanto, este trabalho teve como objetivo relatar os resultados da análise do LCR em 58 ruminantes com distúrbios neurológicos. Amostras do LCR foram obtidas de 32 bovinos, 20 ovinos e 6 caprinos por punção da cisterna cerebelo-medular (n=54) ou espaço lombossacro (n=4) para posterior análise. Esses ruminantes apresentaram sinais neurológicos relacionados a infecções virais (n=13), micóticas (n=3) ou bacterianas (n=15), e desordens tóxicas (n=21), traumáticas (n=4) ou congênitas (n=2) A análise do LCR de ruminantes com infecções virais apresentou pleocitose linfocítica, embora, em vários casos de raiva, o LCR não tenha apresentado alterações. Pleocitose neutrofílica, turbidez, presença de coágulos de fibrina e coloração anormal foram evidentes no LCR da maioria dos casos de infecções bacterianas do SNC, como meningoencefalites, meningites, abscessos, mielite e um caso de conidiobolomicose. Por outro lado, o LCR não foi alterado na maioria dos casos dos distúrbios tóxicos, como botulismo e encefalopatia hepática. A densidade elevada no LCR foi observada em 60% dos ruminantes diagnosticados com polioencefalomalácia. Nossos resultados mostram que a avaliação do LCR é uma valiosa ferramenta de diagnóstico, quando usada em associação com os achados epidemiológicos, clínicos e patológicos para o diagnóstico de doenças do SNC em ruminantes.(AU)