Biomass fractionation

The objective of this master's thesis was to develop a process to increase the value of residual fungal biomass as an animal feed. The increase in value is achieved by enriching the protein content in the biomass and potentially isolating other valuable fractions for productisation. In the literature part of this thesis the composition of fungal biomass and fungal cell wall and the factors affecting them during cultivation are presented. The possible processing options are also presented and evaluated. The soy protein and single cell protein product manufacturing processes are used as examples due to the lack of fungal biomass fractionation processes found in published literature. The second part of this thesis was performed by making laboratory experiments on the developed process, which consisted of acid hydrolysis with subsequent ethanol extraction. Chitin was precipitated from the acid hydrolysate filtrate. The experiments were conducted with three different hydrolysis temperatures and three different acid concentrations. The optimal hydrolysis conditions were 60 °C with 10 %-vol acid concentration. Optimal conditions in hydrolysis resulted in 30 % increase in protein content in the final biomass. The conceptual process was modelled to scale of 10 000 t/a biomass feed. The mass and energy balances were based on the laboratory experiments. Economic calculations were performed to determine the maximal capital expense while achieving 10 % internal rate of return for the investment. For the basic case the capital expense threshold was 25.8 M€. Four optional cases and parameter sensitivity analysis were performed to determine the effects of changes in the process. The chitin sales had the greatest impact of the individual parameters.

Períodos de interferência das plantas daninhas na cultura da soja nos sistemas de semeadura direta e convencional

A presente pesquisa objetivou estudar os efeitos dos períodos de controle e de convivência de uma comunidade infestante sobre as características produtivas da soja nos sistemas de semeadura direta (SSD), com o cultivar CD 201, e convencional (SSC), com a MSOY-6101. No SSD, a comunidade infestante foi composta principalmente por Alternanthera tenella, Cenchrus echinatus e Bidens pilosa, com densidade máxima de 377 plantas m-2, acumulando 365,7 g m-2 de matéria seca, e, no SSC, por A. tenella, Senna obtusifolia e Panicum maximum, com densidade máxima de 161 plantas m-2, acumulando 406,7 g m-2. Tolerando cinco por cento de perda na produtividade da cultura, constatou-se que os períodos críticos de prevenção da interferência foram dos 33 aos 66 DAE para o cultivar CD 201 no SSD e dos 34 aos 76 DAE para o cultivar M-SOY-6101 no SSC. A interferência das plantas daninhas durante todo o ciclo da cultura reduziu, em média, 46% (SSD) e 32% (SSC) a produtividade de grãos da soja. Não houve efeito das plantas daninhas na estatura das plantas, na altura de inserção da primeira vagem e no número de vagens por planta.

Seletividade de diferentes herbicidas à base de glyphosate a soja RR

Com o objetivo de avaliar a seletividade de variedades de soja transgênica (CD 214 RR e M-SOY 8008 RR) a diferentes herbicidas à base de glyphosate (Roundup Ready, Roundup Transorb, Roundup Original, Roundup WG, Polaris, Gliz, Glifosato Nortox e Trop), foi desenvolvido experimento em condições de campo, no ano agrícola 2005/2006, na Fazenda de Ensino, Pesquisa e Produção da UNESP, campus de Jaboticabal, SP. O delineamento experimental utilizado foi o de blocos ao acaso, num esquema de parcelas subdivididas, com quatro repetições. Os herbicidas foram aplicados na dose de 1,2 kg ha-1 de equivalente ácido de glyphosate, quando as plantas de soja se encontravam com a segunda folha trifoliolada totalmente expandida. Não foram observados sintomas de intoxicação dos herbicidas nas plantas de soja. Quanto às demais características avaliadas, a interação variedade x herbicida não foi significativa, indicando que os fatores comportaram-se de forma independente. Entre as variedades, as plantas de M-SOY 8008 RR apresentaram maior altura e número de nós, porém obtiveram menor produtividade de grãos que as de CD 214 RR, devido ao menor número de grãos por vagem. No tocante aos herbicidas, não houve diferença significativa entre eles, ou seja, eles não influenciaram o desenvolvimento vegetativo e reprodutivo das plantas de soja.

Manejo de herbicidas na cultura da soja Roundup Ready®

O trabalho objetivou avaliar a eficácia de diferentes sistemas de manejo de herbicidas no controle de plantas daninhas e no desenvolvimento e produtividade de diferentes cultivares de soja Roundup Ready®. O delineamento experimental utilizado foi o de blocos ao acaso, com quatro repetições, em esquema fatorial 5 x 5, sendo cinco sistemas de aplicação de herbicidas [(1) glyphosate (1.080 g ha-1) + 2,4-D (241,8 g ha-1) 14 dias antes da semeadura (DAS), paraquat + diuron (400 + 200 g ha-1) no dia da semeadura e glyphosate (960 g ha-1) 35 dias após a emergência da cultura (DAE); (2) glyphosate (1.080 g ha-1) + 2,4-D (241,8 g ha-1) aos 14 DAS, paraquat + diuron (400 + 200 g ha-1) no dia da semeadura e glyphosate (480 g ha-1) aos 17 DAE; (3) glyphosate (1.080 g ha-1) + 2,4-D (241,8 g ha-1) aos 14 DAS e glyphosate (960 g ha-1) aos 35 DAE; (4) glyphosate (1.080 g ha-1) + 2,4-D (241,8 g ha-1) aos 14 DAS e glyphosate (480 g ha-1) 17 dias após a emergência das plantas; e (5) testemunha - glyphosate (1.080 g ha-1) + 2,4-D (241,8 g ha-1) aos 14 DAS, sem aplicação de herbicidas em pós-emergência] combinados com cinco cultivares de soja RR® (M-SOY 8585, P98R91, Valiosa, CD 219 e TMG 108), formando 25 tratamentos. Todos os sistemas de aplicação de herbicidas apresentaram controle das espécies de plantas daninhas Chamaesyce hirta, Alternanthera tenella, Euphorbia heterophylla, Spermacoce latifolia e Tridax procumbens superior ao da testemunha sem aplicação de herbicidas em pós-emergência, sendo eficientes no controle dessas espécies. No entanto, no "sistema 3" observou-se menor nível de controle das espécies Spermacoce latifolia, Tridax procumbens e Chamaesyce hirta, esta última somente nas parcelas semeadas com o cultivar CD 219. Os sistemas de manejo de herbicidas influenciaram a altura das plantas de soja, sendo os menores valores verificados no cultivar TMG 108 com a aplicação do "sistema 3". A produtividade de grãos dos cultivares de soja não diferiu entre os sistemas de aplicação de herbicidas, porém todos resultaram em produtividade superior ao da testemunha. O cultivar TMG 108 apresentou maior produtividade de grãos em todos os sistemas de aplicação de herbicidas, inclusive nas parcelas da testemunha, mas não diferindo do cultivar P98R91, nos "sistemas 1, 3 e 4" de aplicação de herbicidas e na testemunha, e do cultivar M-SOY 8585, no "sistema 3".

Eficácia de herbicidas aplicados em pós-emergência na cultura da soja convencional e transgênica

Na cultura da soja é frequente o uso de herbicidas seletivos aplicados em pósemergência para o controle das plantas daninhas. Com o objetivo de avaliar a eficiência e a seletividade de herbicidas aplicados em pós-emergência, isolados e em misturas, para o controle de plantas daninhas e os efeitos sobre o crescimento e o desenvolvimento da soja convencional (M-SOY 8001) e transgênica (M-SOY 7908 RR), foram desenvolvidos dois experimentos em campo, nos anos agrícolas 2006/2007 e 2007/2008. Os tratamentos constaram dos herbicidas: lactofen (168 g ha-1), glyphosate (1.080 g ha-1), lactofen + chlorimuronethyl (96 + 10g ha-1), chlorimuron-ethyl + imazethapyr (10 + 70 g ha-1), chlorimuron-ethyl + bentazon (10 + 600 g ha-1), glyphosate + imazethapyr (900 + 70 g ha-1), lactofen + chlorimuronethyl + imazethapyr (96 + 10 + 70 g ha-1) e lactofen + chlorimuron-ethyl + imazethapyr/ haloxyfop-methyl (96 + 10 + 70 + 60 g ha-1). Além disso, mantiveram-se duas testemunhas sem aplicação de herbicida (capinada e sem capina). O delineamento experimental utilizado foi em blocos ao acaso, com quatro repetições. O herbicida lactofen aplicado isoladamente e em mistura com chlorimuron-etyl, imazethapyr e haloxyfop-methyl provocou intoxicação inicial às plantas de soja, porém aos 27 dias após aplicação a cultura apresentava-se recuperada. No primeiro ano agrícola, as plantas daninhas foram satisfatoriamente controladas pelos tratamentos químicos. No segundo ano, os tratamentos que resultaram em melhor controle das infestantes foram glyphosate e glyphosate + imazethapyr. Houve redução na altura final das plantas, no segundo ano, tratadas com chlorimuron-ethyl + imazethapyr, lactofen + chlorimuron-ethyl + imazethapyr e lactofen + chlorimuron-ethyl + imazethapyr/haloxyfopmethyl. Foi constatada redução na produtividade de grãos da variedade M-SOY 7908 RR tratada com lactofen e com a mistura chlorimuron ethyl + bentazon no primeiro ano. A aplicação de chlorimuron-ethyl + bentazon proporcionou redução na produtividade da variedade M-SOY 8001 no segundo ano.

Resistência de genótipos de soja ao herbicida glyphosate

Avaliou-se a resistência de dez genótipos de soja (Glycine max) ao herbicida glyphosate. Foi adotado o esquema fatorial 10 x 4, no delineamento em blocos casualizados, com quatro repetições. O fator A foi composto por dez genótipos de soja, e o B, por quatro doses de glyphosate. A aplicação do herbicida foi realizada no momento em que as plantas de soja apresentavam a segunda folha trifoliolada completamente desenvolvida. Avaliaramse os caracteres: intoxicação das plantas, número de nós da haste principal, altura das plantas e massa seca das plantas. Na avaliação da intoxicação de plantas, tanto aos quatro dias após a aplicação (DAA) do herbicida quanto aos 28 DAA, os genótipos convencionais apresentaram médias superiores estatisticamente em comparação com genótipos resistentes ao glyphosate (RR). Verificou-se que, nas avaliações realizadas ao 0 DAA ou aos 28 DAA sob 0,0 g e.a. ha¹, as respostas dos genótipos foram diferentes em todos os caracteres avaliados, com exceção do número de nós aos 28 DAA. Essas diferenças podem ser atribuídas aos efeitos fisiológicos e ambientais ou a características do próprio material. Nas demais doses, os genótipos RR comportaram-se de maneira desejável em detrimento dos genótipos convencionais. Ao considerar todos os caracteres avaliados, pode-se afirmar que Valiosa RR, BCR945G110, BCR945G114, BCR892G132, BCR892G140, BCR1067G210, BCR1070G244 e M-SOY 8008RR comportaram-se de forma semelhante quanto à resistência ao glyphosate quando submetidos até a dose de 2.160 g e.a. ha-1 .

Competitiveness of alexandergrass or Bengal dayflower with soybean

Weeds compete with field crops mainly for water, light and nutrients, and the degree of competition is affected by the weed density and the intrinsic competitive ability of each plant species in coexistence. The objective of this research was to compare the competitiveness of alexandergrass (Brachiaria plantaginea) or Bengal dayflower (Commelina benghalensis) in coexistence with soybean, cv. M-Soy 8045. A factorial experiment (2 x 5) with two weed species and five competition proportions was carried out in a completely randomized design with four replicates. Proportions were based on a replacement series competition design, always maintaining the total density of four plants per 10 L plastic pots, which corresponded to 60 plants m ². The weed-crop proportions were: 0:4; 1:3; 2:2; 3:1; 4:0; that corresponded to the proportion of 100, 75, 50, 25 and 0% of soybean plants and the opposite for weeds, B. plantaginea or C. benghalensis plants. Leaf area, shoot dry mass of the weeds and soybean and number of soybean trifoliate leaves were evaluated when the soybean reached the phenologic stage of full flowering. B. plantaginea was a better competitor than soybean plants. Otherwise, C. benghalensis revealed a similar competitive ability that of the soybean. In both cases, there were evidences that intraspecific competition was more important.

Períodos de dessecação de Urochloa ruziziensis e seu reflexo na produtividade da soja RR

Com o objetivo de avaliar os efeitos de períodos de dessecação de Urochloa ruziziensis na implantação e na produtividade da soja transgênica no sistema de plantio direto, foram conduzidos dois experimentos em campo, nos municípios de Colina-SP, no ano agrícola de 2007/08, e Jaboticabal-SP, em 2009/10. Em Colina, utilizou-se uma área de pastagem de U. ruziziensis, conduzida há sete anos. Nesse local foram testados quatro períodos de dessecação da cobertura vegetal: 30, 20, 10 e 0 dias antes da semeadura da soja. No experimento de Jaboticabal, a área utilizada tinha histórico de pastagem com U. ruziziensis, que no ano agrícola 2008/09 foi cultivada com milho. Após a colheita do milho ocorreu reinfestação natural de U. ruziziensis na área, correspondendo a mais de 80% de cobertura vegetal. Nesse experimento, os tratamentos corresponderam a seis períodos de dessecação: 25, 20, 16, 12, 7 e 0 dias após a aplicação. Nos dois experimentos utilizou-se 1,44 kg e.a. ha-1 do herbicida glyphosate e o cultivar de soja M-SOY 7908 RR. O delineamento experimental foi o de blocos ao acaso, com quatro repetições. Foram avaliados o estande da cultura aos 20 dias após a semeadura da soja e na pré-colheita, a altura de plantas, a altura de inserção da primeira vagem, o número de vagens por planta e o rendimento de grãos (produtividade estimada em kg ha-1). Os resultados foram submetidos à análise de variância (teste F), com as médias sendo comparadas pelo teste de Tukey a 5% de probabilidade. A dessecação de U. ruziziensis no mesmo dia da semeadura da soja reduziu a altura das plantas, o número de vagens por planta e a produtividade de grãos. O período recomendado para o manejo químico de U. ruziziensis com glyphosate está entre 10 e 20 dias antes da semeadura da soja. Pôde-se concluir que o período de dessecação de U. ruziziensis alterou a produtividade da cultura.

Density and sowing season of two Brachiaria species on the soybean culture

The aim of this study was to evaluate the growth and yield of soybean cultivar M-8766 in consortium with Brachiaria brizantha. BRS Piata and Brachiaria ruziziensis at different densities and sowing dates. The experimental design was randomized blocks with treatments arranged in a factorial 2 x 2 x 3 with four replications. Used as factors grass species (Brachiaria brizantha Piata and Brachiaria ruziziensis BRS) intercropped with soybean M-8766, sowing dates (12 and 24 days after soybean emergence) and three seeding rates (0, 5, 10 kg ha-1 of seed). At 71 days after soybean emergence were evaluated plant height, stem diameter, dry mass of leaves, stems and shoots, and 4 months after sowing determined the weight of 100 grains and soybean yield. The results showed that when seeded at a density of 10 kg ha-1 at 12 and 24 DAE soy, Brachiaria brizantha. BRS Piata caused reduction in yield in the order of 6.71% and 3.03% respectively, while the Brachiaria ruziziensis was one that caused a greater reduction in productivity in the order of 13.42 and 16.23%, respectively, of these values expression when considering the price of soybean sack. B. ruziziensis expressed less competitive with soybean. However, the large biomass production of this grass provides deployment system till the next harvest.

Interactions between soybean and weeds in a replacement series system, considering the effects of water stress

Due to the increase of water deficiency in many farm regions and its meaning on weed interference, competitive interactions between soybean and three weeds were evaluated under water stress (20 to 40 days after transplanting) and no stress conditions. Three independent experiments were carried out in a growth chamber, being each one composed by the weeds Alternanthera tenella, Tridax procumbens or Digitaria ciliaris, along with the crop, in which soil water condition and plant composition effects were evaluated while in competition. A replacement series system was used, including both monoculture of each species and a mixture with a ratio of 50% between weed and soybean. A completely randomized design was used in factorial arrangement, with treatments distributed in three levels for plant composition factor (soybean and weeds monocultures, in addition to the soybean + weed mixture) and two levels for the water factor (with or without stress), amounting six treatments in each experiment. Soybean dry mass was higher than weed dry mass, when growing without water stress. However, under water stress conditions, the dry mass of soy was reduced in all experiments, mainly in the D. ciliaris comparative experiment. Water restriction was also significant in the plants' photosynthesis reduction in most of the experiments, reducing leaf area duration and efficiency of water use. Analysing all variables shows greater weed tolerance than soybean when submitted to water deficit and with distinct changes of their interactions and mechanism of competition, in each experiment.

Behavior of RR soybeans subjected to different formulations and rates of glyphosate in the reproductive period

Recent studies indicate that glyphosate applied in post-emergence in RR soybean can eventually cause phytotoxic effects. However, there are many questions that need to be clarified in the scientific and technical contexts, involving the issue of RR soybeans regarding the use of glyphosate. This study has assessed the impact of the application of different doses and formulations of glyphosate in the reproductive period of RR soybean (R1 stage). For that purpose, an experiment in the field was conducted in two harvests (2011/12 and 2012/13), in which a 2 x 5 factorial design was used (formulations versus doses) totaling 10 treatments. In these two experiments the variables related to agronomic performance were: phytotoxicity (7, 14, 21 and 28 days after application), plant height, number of pods per plant, yield and weight of 100 grains (end of soy cycle). The results obtained allowed characterizing phytotoxicity and damages to the height and yield in RR soybean, with increasing rates of glyphosate applied in the reproductive period.

Apolipoprotein and lipid abnormalities in chronic liver failure

Total serum lipids, as well as apolipoproteins A-I (apo A-I) and B (apo B), were determined in 74 patients with chronic liver failure without cholestasis and in 82 normal subjects. The VLDL, LDL and HDL lipid fractions were reduced in the liver failure group by 36%, 24% and 46%, respectively (P<0.001). Apolipoproteins A-I and B were also reduced by 26% and 25%, respectively (P<0.001). However, the reduction of HDL cholesterol (HDLc) was more pronounced than that of apo A-I and the HDLc:apo A-I ratio was significantly lower in the liver failure group. After separating these patients into groups with plasma albumin lower than 3.0, between 3.0 and 3.5, and higher than 3.5 g/dl, the HDLc:apo A-I ratio was proportional to plasma albumin, but the correlation was not statistically significant. When these patients were separated by the Child classification of liver function, there was a correlation between the HDLc:apo A-I ratio and liver function. The differences in the HDLc:apo A-I ratio between the Child groups B and C, and A and C were statistically significant (P<0.05). We conclude that there is a more pronounced reduction in HDL cholesterol than in apo A-I in liver failure patients. Therefore, the HDLc:apo A-I ratio is a marker of liver function, probably because there is a decreased lecithin-cholesterol acyltransferase production by the diseased liver

Preparation and purification of Flavobacterium heparinum chondroitinases AC and B by hydrophobic interaction chromatography

Flavobacterium heparinum is a soil bacterium that produces several mucopolysaccharidases such as heparinase, heparitinases I and II, and chondroitinases AC, B, C and ABC. The purpose of the present study was to optimize the preparation of F. heparinum chondroitinases, which are very useful tools for the identification and structural characterization of chondroitin and dermatan sulfates. We observed that during the routine procedure for cell disruption (ultrasound, 100 kHz, 5 min) some of the chondroitinase B activity was lost. Using milder conditions (2 min), most of the chondroitinase B and AC protein was solubilized and the enzyme activities were preserved. Tryptic soy broth without glucose was the best culture medium both for bacterial growth and enzyme induction. Chondroitinases AC and B were separated from each other and also from glucuronidases and sulfatases by hydrophobic interaction chromatography on HP Phenyl-Sepharose. A rapid method for screening of the column fractions was also developed based on the metachromatic shift of the color of dimethylmethylene blue.

Neuroprotection by flavonoids

The high morbidity, high socioeconomic costs and lack of specific treatments are key factors that define the relevance of brain pathology for human health and the importance of research on neuronal protective agents. Epidemiological studies have shown beneficial effects of flavonoids on arteriosclerosis-related pathology in general and neurodegeneration in particular. Flavonoids can protect the brain by their ability to modulate intracellular signals promoting cellular survival. Quercetin and structurally related flavonoids (myricetin, fisetin, luteolin) showed a marked cytoprotective capacity in in vitro experimental conditions in models of predominantly apoptotic death such as that induced by medium concentrations (200 µM) of H2O2 added to PC12 cells in culture. Nevertheless, quercetin did not protect substantia nigra neurons in vivo from an oxidative insult (6-hydroxydopamine), probably due to difficulties in crossing the blood-brain barrier. On the other hand, treatment of permanent focal ischemia with a lecithin/quercetin preparation decreased lesion volume, showing that preparations that help to cross the blood-brain barrier may be critical for the expression of the effects of flavonoids on the brain. The hypothesis is advanced that a group of quercetin-related flavonoids could become lead molecules for the development of neuroprotective compounds with multitarget anti-ischemic effects.

Atherosclerosis in aged mice over-expressing the reverse cholesterol transport genes

We determined whether over-expression of one of the three genes involved in reverse cholesterol transport, apolipoprotein (apo) AI, lecithin-cholesterol acyl transferase (LCAT) and cholesteryl ester transfer protein (CETP), or of their combinations influenced the development of diet-induced atherosclerosis. Eight genotypic groups of mice were studied (AI, LCAT, CETP, LCAT/AI, CETP/AI, LCAT/CETP, LCAT/AI/CETP, and non-transgenic) after four months on an atherogenic diet. The extent of atherosclerosis was assessed by morphometric analysis of lipid-stained areas in the aortic roots. The relative influence (R²) of genotype, sex, total cholesterol, and its main sub-fraction levels on atherosclerotic lesion size was determined by multiple linear regression analysis. Whereas apo AI (R² = 0.22, P < 0.001) and CETP (R² = 0.13, P < 0.01) expression reduced lesion size, the LCAT (R² = 0.16, P < 0.005) and LCAT/AI (R² = 0.13, P < 0.003) genotypes had the opposite effect. Logistic regression analysis revealed that the risk of developing atherosclerotic lesions greater than the 50th percentile was 4.3-fold lower for the apo AI transgenic mice than for non-transgenic mice, and was 3.0-fold lower for male than for female mice. These results show that apo AI overexpression decreased the risk of developing large atherosclerotic lesions but was not sufficient to reduce the atherogenic effect of LCAT when both transgenes were co-expressed. On the other hand, CETP expression was sufficient to eliminate the deleterious effect of LCAT and LCAT/AI overexpression. Therefore, increasing each step of the reverse cholesterol transport per se does not necessarily imply protection against atherosclerosis while CETP expression can change specific athero genic scenarios.