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In Brazil the main feedstock used for ethanol production is sugarcane juice, resulting in large amounts of bagasse. Bagasse has high potential for cellulosic ethanol production, and consolidated bioprocessing (CBP) has potential for lowering costs. However, economic feasibility requires bioprocessing at high solids loadings, entailing engineering and biological challenges. This study aims to document and characterize carbohydrate solubilization and utilization by defined cocultures of Clostridium thermocellum and Thermoanaerobacterium thermosaccharolyticum at increasing loadings of sugarcane bagasse. Results show that fractional carbohydrate solubilization decreases as solids loading increases from 10 g/L to 80 g/L. Cocultures enhance solubilization and carbohydrate utilization compared to monocultures, irrespective of initial solids loading. Rinsing bagasse before fermentation slightly decreases solubilization. Experiments studying inhibitory effects using spent media and dilution of broth show that negative effects are temporary or reversible. These findings highlight the potential of converting sugarcane bagasse via CBP, pointing out performance limitations that must be addressed.

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Cotreatment, mechanical disruption of lignocellulosic biomass during microbial fermentation, is a potential alternative to thermochemical pretreatment as a means of increasing the accessibility of lignocellulose to biological attack. Successful implementation of cotreatment requires microbes that can withstand milling, while solubilizing and utilizing carbohydrates from lignocellulose. In this context, cotreatment with thermophilic, lignocellulose-fermenting bacteria has been successfully evaluated for a number of lignocellulosic feedstocks. Here, cotreatment was applied to sugarcane bagasse using monocultures of the cellulose-fermenting Clostridium thermocellum and cocultures with the hemicellulose-fermenting Thermoanaerobacterium thermosaccharolyticum. This resulted in 76 % carbohydrate solubilization (a 1.8-fold increase over non-cotreated controls) on 10 g/L solids loading, having greater effect on the hemicellulose fraction. With cotreatment, fermentation by wild-type cultures at low substrate concentrations increased cumulative product formation by 45 % for the monoculture and 32 % for the coculture. These findings highlight the potential of cotreatment for enhancing deconstruction of sugarcane bagasse using thermophilic bacteria.

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Biodegradable starch foam trays offer an eco-friendly substitute for petroleum-based single-use packaging, notably polystyrene foams. However, they lack flexibility, tensile strength, and water-sensitivity, addressable through lignocellulosic reinforcement. This study aimed to develop biodegradable starch foam trays filled with different food-chain side streams for sustainable alternative packaging. Corncob, soybean straw, cassava peel, araucaria seed hull, yerba mate stalks and yerba mate leaves petiole were collected, dried and ground to <250 µm. The trays were filled with 13 % (w/w) of each food-chain side streams and produced by hot molding. The trays morphology, moisture, water activity (aw), thickness, bulk density, tensile strength, elongation at break, Young's modulus, bending strength, maximum deflection, and sorption isotherms were investigated. Reinforcements slightly increased the foams bulk density, reduced the tensile strength and maximum deflection and while bending strength increased from 0.20 MPa to 1.17-1.80 MPa. The elasticity modulus decreased by adding any filling, that resulted in ductility improvement; however, these packaging have moisture-sensitive material especially for aw higher than 0.52, which drives the use recommendation for dry products storage or shipping/transport. The biodegradable starch foam trays filled with side streams were successfully produced and offer excellent alternative to petroleum-based packaging low-density material with bending strength improved.

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Trichoderma erinaceum is a filamentous fungus that was isolated from decaying sugarcane straw at a Brazilian ethanol biorefinery. This fungus shows potential as a source of plant cell wall-degrading enzymes (PCWDEs). In this study, we conducted a comprehensive multiomics investigation of T. erinaceum to gain insights into its enzymatic capabilities and genetic makeup. Firstly, we performed genome sequencing and assembly, which resulted in the identification of 10,942 genes in the T. erinaceum genome. We then conducted transcriptomics and secretome analyses to map the gene expression patterns and identify the enzymes produced by T. erinaceum in the presence of different substrates such as glucose, microcrystalline cellulose, pretreated sugarcane straw, and pretreated energy cane bagasse. Our analyses revealed that T. erinaceum highly expresses genes directly related to lignocellulose degradation when grown on pretreated energy cane and sugarcane substrates. Furthermore, our secretome analysis identified 35 carbohydrate-active enzymes, primarily PCWDEs. To further explore the enzymatic capabilities of T. erinaceum, we selected a ß-glucosidase from the secretome data for recombinant production in a fungal strain. The recombinant enzyme demonstrated superior performance in degrading cellobiose and laminaribiose compared to a well-known enzyme derived from Trichoderma reesei. Overall, this comprehensive study provides valuable insights into both the genetic patterns of T. erinaceum and its potential for lignocellulose degradation and enzyme production. The obtained genomic data can serve as an important resource for future genetic engineering efforts aimed at optimizing enzyme production from this fungus.

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Trametes villosa is a remarkable white-rot fungus (WRF) with the potential to be applied in lignocellulose conversion to obtain chemical compounds and biofuels. Lignocellulose breakdown by WRF is carried out through the secretion of oxidative and hydrolytic enzymes. Despite the existing knowledge about this process, the complete molecular mechanisms involved in the regulation of this metabolic system have not yet been elucidated. Therefore, in order to understand the genes and metabolic pathways regulated during lignocellulose degradation, the strain T. villosa CCMB561 was cultured in media with different carbon sources (lignin, sugarcane bagasse, and malt extract). Subsequently, biochemical assays and differential gene expression analysis by qPCR and high-throughput RNA sequencing were carried out. Our results revealed the ability of T. villosa CCMB561 to grow on lignin (AL medium) as the unique carbon source. An overexpression of Cytochrome P450 was detected in this medium, which may be associated with the lignin O-demethylation pathway. Clusters of up-regulated CAZymes-encoding genes were identified in lignin and sugarcane bagasse, revealing that T. villosa CCMB561 acts simultaneously in the depolymerization of lignin, cellulose, hemicellulose, and pectin. Furthermore, genes encoding nitroreductases and homogentisate-1,2-dioxygenase that act in the degradation of organic pollutants were up-regulated in the lignin medium. Altogether, these findings provide new insights into the mechanisms of lignocellulose degradation by T. villosa and confirm the ability of this fungal species to be applied in biorefineries and in the bioremediation of organic pollutants.

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The lignocellulose bioconversion process is an eco-friendly and green-economy alternative technology that allows the reduction of pollution and global warming, so it is necessary for thermophilic and thermostable hydrolytic enzymes from natural sources. This research aimed to isolate cellulolytic and xylanolytic microbial consortia from Huancarhuaz hot spring (Peru) from sludge or in situ baiting cultured with or without sugarcane bagasse. According to the hydrolytic activities consortium T4 from in situ baiting was selected. It was cultivated in submerged fermentation at 65 °C, pH 6.5 for eight days using LB supplemented with sugar cane bagasse (SCB), pine wood sawdust (PWS), CMC, xylan of birchwood, or micro granular cellulose. Crude extract of culture supplemented with SCB (T4B) showed better endoglucanase and xylanase activities with higher activities at 75 °C and pH 6. In these conditions, cellulase activity was kept up to 57% after 1 h of incubation, while xylanase activity was up to 63% after 72 h. Furthermore, this crude extract released reduced sugars from pretreated SCB and PWS. According to metagenomic analysis of 16S rDNA, Geobacillus was the predominant genus. It was found thermostable genes: a type of endoglucanase (GH5), an endo-xylanase (GH10), and alkali xylanase (GH10) previously reported in Geobacillus sp. strains. Finally, Huancarhuaz hot spring harbors a genetic microbial diversity for lignocellulosic waste bioconversion in high temperatures, and the T4B consortium will be a promising source of novel extreme condition stable enzymes for the saccharification process.

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The agricultural sugarcane residues, bagasse and straws, can be used for second-generation ethanol (2GE) production by the cellulose conversion into glucose (saccharification). However, the lignin content negatively impacts the saccharification process. This polymer is mainly composed of guaiacyl (G), hydroxyphenyl (H), and syringyl (S) units, the latter formed in the ferulate 5-hydroxylase (F5H) branch of the lignin biosynthesis pathway. We have generated transgenic lines overexpressing ShF5H1 under the control of the C4H (cinnamate 4-hydroxylase) rice promoter, which led to a significant increase of up to 160% in the S/G ratio and 63% in the saccharification efficiency in leaves. Nevertheless, the content of lignin was unchanged in this organ. In culms, neither the S/G ratio nor sucrose accumulation was altered, suggesting that ShF5H1 overexpression would not affect first-generation ethanol production. Interestingly, the bagasse showed a significantly higher fiber content. Our results indicate that the tissue-specific manipulation of the biosynthetic branch leading to S unit formation is industrially advantageous and has established a foundation for further studies aiming at refining lignin modifications. Thus, the ShF5H1 overexpression in sugarcane emerges as an efficient strategy to improve 2GE production from straw.

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Two trials were performed in order to evaluate the effects of dietary Kraft lignin inclusion on broiler performance, ileal nutrient digestibility, blood lipid profile, intestinal morphometry, and lipid oxidation of meat. Trial 1 was conducted in order to evaluate performance and ileal digestibility for the period of 1 to 21 d of age, randomly distributing 490 day-old broiler chicks across 5 dietary treatments with 14 replicates containing 7 birds each in metabolic cages, while trial 2 was executed in order to evaluate performance, blood parameters, intestinal morphometry, carcass yield and abdominal fat, and lipid oxidation for the period of 1 to 42 d of age, randomly distributing 900 day-old broiler chicks across 5 dietary treatments with 15 replicates of 12 birds each in floor pens, being each bird in trial 2 challenged with coccidiosis vaccine at 10 d of age. The treatments used in both trials were: positive control (PC): basal diet + antimicrobial; negative control (NC): Basal diet; NC1: NC + 1% lignin; NC2: NC + 2% lignin; NC3: NC + 3% lignin. For trial 1, it was observed that birds fed diets containing 1% lignin had a significant positive effect for BW, feed intake (FI), average daily weight gain (BWG) and feed conversion rate (FCR), similar to the PC, but also showing better EE, CP and AAs ileal digestibility percentages when compared to other treatments. For trial 2, it was observed that during the period of 21 to 35 d, the inclusion of lignin in the diet provided better results in animal performance, similar to the PC group, but at 42 d, animals fed with dietary lignin showed results lower than animals fed the PC diet (P < 0.05). Animals fed with increasing lignin concentrations showed decreasing levels of HDL (P < 0.05). As of intestinal morphometry, animals fed with 1% and 3% lignin showed longer intestinal length (P < 0.05). At 14 d of age, it was observed that animals fed with lignin showed oxidation levels similar to the control treatments. The inclusion of up to 1% lignin in the diet provides beneficial effects on productive performance and nutrient digestibility, while the inclusion of 2% lignin provided lower cholesterol levels, lower villus/crypt ratio, and better internal organ development, therefore, it can be considered an alternative to performance-enhancing antimicrobials in broiler chicken diets.

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Radiation chemistry presents a unique avenue for developing innovative polymeric materials with desirable properties, eliminating the need for chemical initiators, which can be potentially detrimental, especially in sensitive sectors like medicine. In this investigation, we employed a radiation-induced graft polymerization process with N-vinylcaprolactam (NVCL) to modify lignocellulosic membranes derived from Agave salmiana, commonly known as maguey. The membranes underwent thorough characterization employing diverse techniques, including contact angle measurement, degree of swelling, scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier-transform infrared-attenuated total reflectance spectroscopy (FTIR-ATR), nuclear magnetic resonance (CP-MAS 13C-NMR), X-ray photoelectron spectroscopy (XPS), and uniaxial tensile mechanical tests. The membranes' ability to load and release an antimicrobial glycopeptide drug was assessed, revealing significant enhancements in both drug loading and sustained release. The grafting of PNVCL contributed to prolonged sustained release by decreasing the drug release rate at temperatures above the LCST. The release profiles were analyzed using the Higuchi, Peppas-Sahlin, and Korsmeyer-Peppas models, suggesting a Fickian transport mechanism as indicated by the Korsmeyer-Peppas model.

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This study evaluated the effect of diets containing low levels of soluble and insoluble fiber sources on performance, diarrhea score, chemical and physical characteristics of feces, and behavior in weaning piglets. Thirty-six weaned piglets of 30 days of age with an initial body weight of 8.8 kg were distributed in 36 pens using a completely randomized design in an experimental period of 21 days. The experimental treatments were the Control diet (CONT), the Control diet + 1% beet pulp inclusion (SBP), and the Control diet + 1% lignocellulose inclusion (LCE, Arbocel®). Feed and water were available ad libitum. Body weight and feed intake were measured weekly to calculate the average daily intake, weight gain, and feed conversion ratio. The fecal consistency was determined visually twice daily, classifying feces according to three scores. To determine fecal pH and concentration of fecal short-chain fatty acids (SCFA), samples of fresh feces were collected two weeks after weaning and measured by a digital pH meter and gas chromatography, respectively. The behavior of piglets was observed once a week, using four animals per treatment, from 14:00 to 16:00, every 12 min. Fibre sources had no effect (P>0.05) on performance, except in the period 15 to 21 days after weaning, which was a tendency (P=0.061) of feed intake decrease in SBP and LCE diets. Fiber sources did not affect the fecal consistency score (P>0.05). However, piglets fed SBP and LCE showed a tendency (P<0.10) to have less diarrhea incidence 15 to 21 days post-weaning and in the entire experimental period. Fecal pH and SCFA concentration were not influenced by fiber source (P>0.05), with acetic, propionic, and butyric acids representing around 71%, 19%, and 10% of the total, respectively. Fiber sources did not influence the social and feeding behavior of weaning piglets (P>0.05). Diets containing 1% fiber sources did not alter performance, diarrhea score, fecal pH, fecal SCFA concentration, or feeding and social behavior of weaned piglets.(AU)

O estudo avaliou o efeito de dietas contendo baixos níveis de fontes de fibra solúvel e insolúvel sobre o desempenho, escore de diarreia, características químicas e físicas das fezes e comportamento de leitões desmamados. Trinta e seis leitões desmamados, com 30 dias de idade e peso vivo inicial de 8,8 kg, foram distribuídos em 36 baias, totalizando 12 repetições por tratamento, em um delineamento inteiramente casualizado. Os tratamentos experimentais foram: dieta controle (CONT), dieta controle + 1% de inclusão de polpa de beterraba (SBP) e dieta controle + 1% de inclusão de lignocelulose (LCE, Arbocel®). A ração e a água foram disponibilizadas ad libitum durante os 21 dias experimentais. O consumo médio diário de ração (CRM), ganho de peso diário (GPD) e a conversão alimentar (CA) foram medidos semanalmente. A consistência fecal foi determinada visualmente duas vezes por dia, classificando as fezes de acordo com três classificações. Amostras frescas de fezes, colhidas no 14° dia experimental, foram usadas para determinação do pH e ácidos graxos de cadeia curta (AGCC). O pH foi medido utilizando pHmetro digital, enquanto os AGCC foram determinados com auxílio de cromatografia gasosa. Para avaliar o comportamento foram observados quatro leitões por tratamento, uma vez por semana, das 14:00 às 16:00, a cada 12 minutos. As fontes de fibra não apresentaram efeito (P>0,05) sobre as variáveis de desempenho, exceto no período de 15 a 21 dias pós desmame, onde se observou uma tendência (P=0,061) de redução no consumo médio nos leitões que receberam as dietas contendo SBP e LCE. Não foi observado efeito das fontes de fibra sobre o escore de consistência fecal (P>0,05), embora leitões alimentados com SBP e LCE apresentaram uma tendência (P<0,10) de redução na incidência de diarreia no período de 15 a 21 dias pós desmame e no período total. O pH e a concentração de AGCC não foram influenciados pelas fontes de fibra (P>0,05), onde entres os tratamentos os perfis dos ácidos acético, propiônico e butírico foram semelhantes 71%, 19% e 10%, respectivamente. Não houve efeito das fontes de fibra sobre os comportamentos social e alimentar dos leitões (P>0,05). Dietas contendo 1% de fontes de fibra não alteram o desempenho, escore de diarreia, pH fecal, concentração de ácidos graxos voláteis nas fezes, bem como o comportamento alimentar e social de leitões desmamados.(AU)

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