Fermentação ruminal e produção de metano em bovinos alimentados com feno de capim-tifton 85 e concentrado com aditivos

Objetivou-se avaliar o efeito do uso de monensina, complexo de leveduras, ácidos graxos poliinsaturados e aminoácidos no consumo de matéria seca e nutrientes, na estimativa da digestibilidade ruminal, nos parâmetros de fermentação ruminal (pH, concentração de nitrogênio amoniacal e de ácidos graxos de cadeia curta), na população de protozoários e na produção de metano. Foram utilizados seis bovinos e com peso corporal de 530 ± 15 kg, recebendo complexo de leveduras, ácidos graxos poliinsaturados e aminoácidos (5 g/dia); monensina (5 g/dia); caulim (5 g/dia), usado como controle adicionado à dieta composta de feno de capim-tifton 85 (Cynodon spp.); e concentrado, na relação 80:20. O delineamento experimental adotado para análise do consumo e da digestibilidade foi o de blocos completos casualizados e, para análise dos parâmetros ruminais e da produção de metano, o de parcelas subdivididas. O consumo foi influenciado pelo uso de monensina na dieta, mas não diferiu entre os aditivos. As digestibilidades da matéria seca e dos nutrientes não foram influenciadas pelo fornecimento dos aditivos. A relação acetato:propionato nos animais alimentados com a dieta com monensina foi menor que naqueles que receberam o complexo de leveduras e ácidos graxos poliinsaturados e aminoácidos, diminuindo a perda de energia na forma de metano. O pH e a concentração de nitrogênio amoniacal foram adequados para o crescimento bacteriano. A concentração de metano não é alterada pelo uso dos aditivos testados.

Fermentação alcoólica utilizando líquido da casca de coco verde como fonte de nutrientes

The liquid of the rind of green coconut (LCCV), an effluent stream from the industrial processing of green coconut rind, is rich in sugars and is a suitable feedstock for fermentation. The first step of this study was to evaluate the potential of natural fermentation of LCCV. As the literature did not provide any information about LCCV and due to the difficulty of working with such an organic effluent, the second step was to characterize the LCCV and to develop a synthetic medium to explore its potential as a bioprocess diluent. The third step was to evaluate the influence of initial condensed and hydrolysable tannins on alcoholic fermentation. The last step of this work was divided into several stages: in particular to evaluate (1) the influence of the inoculum, temperature and agitation on the fermentation process, (2) the carbon source and the use of LCCV as diluent, (3) the differences between natural and synthetic fermentation of LCCV, in order to determine the best process conditions. Characterization of LCCV included analyses of the physico-chemical properties as well as the content of DQO, DBO and series of solids. Fermentation was carried out in bench-scale bioreactors using Saccharomyces cerevisiae as inoculum, at a working volume of 5L and using 0.30% of soy oil as antifoam. During fermentations, the effects of different initial sugars concentrations (10 - 20%), yeast concentrations (5 and 7.5%), temperatures (30 - 50°C) and agitation rates (400 and 500 rpm) on pH/sugars profiles and ethanol production were evaluated. The characterization of LCCV demonstrated the complexity and variability of the liquid. The best conditions for ethanol conversion were (1) media containing 15% of sugar; (2) 7.5% yeast inoculum; (3) temperature set point of 40°C and (4) an agitation rate of 500 rpm, which resulted in an ethanol conversion rate of 98% after 6 hours of process. A statistical comparison of results from natural and synthetic fermentation of LCCV showed that both processes are similar

Efeito de variáveis de processo no tempo de fermentação e na concentração das dicetonas vicinais totais

Among the main challenges in the beer industrial production is the market supply at the lowest cost and high quality, in order to ensure the expectations of customers and. consumers The beer fermentation stage represents approximately 70% of the whole time necessary to its production, having a obligatoriness of strict process controls to avoid becoming bottleneck in beer production. This stage is responsible for the formation of a series of subproducts, which are responsible for the composition of aroma/bouquet existing in beer and some of these subproducts, if produced in larger quantities, they will confer unpleasant taste and odor to the final product. Among the subproducts formed during the fermentation stage, total vicinal diketones is the main component, since it is limiting for product transfusion to the subsequent steps, besides having a low perception threshold by the consumer and giving undesirable taste and odor. Due to the instability of main raw materials quality and also process controls during fermentation, the development of alternative forms of beer production without impacting on total fermentation time and final product quality is a great challenge to breweries. In this work, a prior acidification of the pasty yeast was carried out, utilizing for that phosphoric acid, food grade, reducing yeast pH of about 5.30 to 2.20 and altering its characteristic from flocculent to pulverulent during beer fermentation. An increase of six times was observed in amount of yeast cells in suspension in the second fermentation stage regarding to fermentations by yeast with no prior acidification. With alteration on two input variables, temperature curve and cell multiplication, which goal was to minimize the maximum values for diketones detected in the fermenter tank, a reduction was obtained from peak of formed diacetyl and consequently contributed to reduction in fermentation time and total process time. Several experiments were performed with those process changes in order to verify the influence on the total fermentation time and total vicinal diketones concentration at the end of fermentation. This experiment reached as the best production result a total fermentation time of 151 hours and total vicinal diketone concentration of 0.08 ppm. The mass of yeast in suspension in the second phase of fermentation increased from 2.45 x 106 to 16.38 x 106 cells/mL of yeast, which fact is key to a greater efficiency in reducing total vicinal diketones existing in the medium, confirming that the prior yeast acidification, as well as the control of temperature and yeast cell multiplication in fermentative process enhances the performance of diketones reduction and consequently reduce the total fermentation time with diketones concentration below the expected value (Max: 0.10 ppm)

Produção de enzimas por fungos em fermentação semi-sólida utilizando bagaço de coco e pedúnculo de caju como substratos

The production of enzymes by microorganisms using organic residues is important and it can be associated with several applications such as food and chemical industries and so on. The objective of this work is the production of CMCase, Xylanase, Avicelase and FPase enzymes by solid state fermentation (SSF) using as substrates: bagasse of coconut and dried cashew stem. The microorganisms employed are Penicillium chrysogenum and an isolated fungus from the coconut bark (Aspergillus fumigatus). Through the factorial design methodology and response surface analysis it was possible to study the influence of the humidity and pH. For Penicillium chrysogenum and the isolated fungus, the coconut bagasse was used as culture medium. In another fermentation, it was used the mixture of coconut bagasse and cashew stem. Fermentations were conducted using only the coconut bagasse as substrate in cultures with Penicillium chrysogenum fungus and the isolated one. A mixture with 50% of coconut and 50% of cashew stem was employed only for Penicillium chrysogenum fungus, the cultivation conditions were: 120 hours at 30 °C in BOD, changing humidity and pH values. In order to check the influence of the variables: humidity and pH, a 2 2 factorial experimental design was developed, and then two factors with two levels for each factor and three repetitions at the central point. The levels of the independent variables used in ascending order (-1, 0, +1), to humidity, 66%, 70.5% and 75% and pH 3, 5 and 7, respectively. The software STATISTICA TM (version 7.0, StatSoft, Inc.) was used to calculate the main effects of the variables and their interactions. The response surface methodology was used to optimize the conditions of the SSF. A chemical and a physic-chemical characterization of the coconut bagasse have determined the composition of cellulose (%) = 39.09; Hemicellulose (%) = 23.80, Total Lignin (%) = 36.22 and Pectin (%) = 1.64. To the characterization of cashew stem, the values were cellulose (g) = 15.91 Hemicellulose (%) = 16.77, Total Lignin (%) = 30.04 and Pectin (%) = 15.24. The results indicate the potential of the materials as substrate for semisolid fermentation enzyme production. The two microorganisms used are presented as good producers of cellulases. The results showed the potential of the fungus in the production of CMCase enzyme, with a maximum of 0.282 UI/mL and the Avicelase enzyme the maximum value ranged from 0.018 to 0.020 UI/ mL, using only coconut bagasse as substrate. The Penicillium chrysogenum fungus has showed the best results for CMCase = 0.294 UI/mL, FPase = 0.058 UI/mL, Avicelase = 0.010 UI/mL and Xylanase = 0.644 UI/ mL enzyme, using coconut bagasse and cashew stem as substrates. The Penicllium chrysogenum fungus showed enzymatic activities using only the coconut as substrate for CMCase = 0.233 UI/mL, FPase = 0.031 to 0.032 UI/ mL, Avicelase = 0.018 to 0.020 UI/mL and Xylanase = 0.735 UI/ mL. Thus, it can be concluded that the used organisms and substrates have offered potential for enzyme production processes in a semi-solid cultivation

Estudo da produção de pectinase por fermentação em estado sólido utilizando pedúnculo de caju como substrato

Pectinolytic enzymes, or simply pectinases, are complex enzymes that degrade pectic polymers. They have many uses, such as fruit juice extraction and purification, textile fiber treatment and vegetal oil extraction. The aim of this work was to study the kinetics of pectinases production by solid-state fermentation, using dry cashew apple residue as substrate and the microorganism Aspergillus niger CCT 0916. The influence of the initial medium moisture and medium supplementation with a source of nitrogen and phosphorus was evaluated using the factorial experimental planning and response surface methodology. Ammonia sulphate and potassium phosphate were used as nitrogen and phosphorus source, respectively. The variables time of contact (T) and ratio volume solvent/fermented medium (RZ), in systems with and without agitation, were evaluated in order to study the best extraction condition of the produced enzyme. Washed and unwashed cashew apple residues were tested as the growth medium. The unwashed residue was obtained by drying the residue after the extraction of the juice, while the washed residue was obtained by water washing 5 times using the proportion of 1 kg pulp/2 liters of water. Samples were taken every 12 hours for moisture content, pH, protein, reducing sugars, polygalacturonase activity (PG) and viscosity reduction. The physical-chemical composition of the residues had different sugar and pectin levels. For the unwashed residue, the peak activity was reached with 40% of initial moisture content, 1% of nitrogen supplementation without phosphorus addition after 30 hours of process. These conditions led to 16 U/g of PG activity and 82% of viscosity reduction. The calculated models reached similar values to the experimental ones in the same process conditions: 15.55 U/g of PG and 79.57% of viscosity eduction. Similarly, the greatest enzyme production for washed residue was reached with 40% initial moisture content, 1% nitrogen supplementation without phosphorus addition after 22 hours of cultivation. In this condition it was obtained polygalacturonase activity of 9.84 U/g and viscosity reduction of 81.36%. These values are close to experimental values that were of 10.1 U/g and 81%, respectively. The conditions that led to the best PG activity results was the agitated one and the best extraction condition was obtained with 100 minutes of solvent/medium contact and RZ of 5 (mL/g)

Produção de bioetanol a partir de pedúnculo de caju (Anacardium occidentale L.) por fermentação submersa

Recently, global demand for ethanol fuel has expanded very rapidly, and this should further increase in the near future, almost all ethanol fuel is produced by fermentation of sucrose or glucose in Brazil and produced by corn in the USA, but these raw materials will not be enough to satisfy international demand. The aim of this work was studied the ethanol production from cashew apple juice. A commercial strain of Saccharomyces cerevisiae was used for the production of ethanol by fermentation of cashew apple juice. Growth kinetics and ethanol productivity were calculated for batch fermentation with different initial sugar (glucose + fructose) concentration (from 24.4 to 103.1 g.L-1). Maximal ethanol, cell and glycerol concentrations (44.4 g.L-1, 17.17 g.L-1, 6.4 g.L-1, respectively) were obtained when 103.1 g.L-1 of initial sugar concentration were used, respectively. Ethanol yield (YP/S) was calculated as 0.49 g (g glucose + fructose)-1. Pretreatment of cashew apple bagasse (CAB) with dilute sulfuric acid was investigated and evaluated some factors such as sulfuric acid concentration, solid concentration and time of pretreatment at 121°C. The maximum glucose yield (162.9 mg/gCAB) was obtained by the hydrolysis with H2SO4 0.6 mol.L-1 at 121°C for 15 min. Hydrolysate, containing 16 ± 2.0 g.L-1 of glucose, was used as fermentation medium for ethanol production by S. cerevisiae and obtained a ethanol concentration of 10.0 g.L-1 after 4 with a yield and productivity of 0.48 g (g glucose)-1 and 1.43 g.L-1.h-1, respectively. The enzymatic hydrolysis of cashew apple bagasse treated with diluted acid (CAB-H) and alkali (CAB-OH) was studied and to evaluate its fermentation to ethanol using S. cerevisiae. Glucose conversion of 82 ± 2 mg per g CAB-H and 730 ± 20 mg per g CAB-OH was obtained when was used 2% (w/v) of solid and loading enzymatic of 30 FPU/g bagasse at 45 °C. Ethanol concentration and productivity was achieved of 20.0 ± 0.2 g.L-1 and 3.33 g.L-1.h-1, respectively when using CAB-OH hydrolyzate (initial glucose concentration of 52.4 g.L-1). For CAB-H hydrolyzate (initial glucose concentration of 17.4 g.L-1), ethanol concentration and productivity was 8.2 ± 0.1 g.L-1 and 2.7 g.L-1.h-1, respectively. Hydrolyzates fermentation resulted in an ethanol yield of 0.38 g/g glucose and 0.47 g/g glucose, with pretreated CABOH and CAB-H, respectively. The potential of cashew apple bagasse as a source of sugars for ethanol production by Kluyveromyces marxianus CE025 was evaluated too in this work. First, the yeast CE025 was preliminary cultivated in a synthetic medium containing glucose and xylose. Results showed that it was able to produce ethanol and xylitol at pH 4.5. Next, cashew apple bagasse hydrolysate (CABH) was prepared by a diluted sulfuric acid pre-treatment. The fermentation of CABH was conducted at pH 4.5 in a batch-reactor, and only ethanol was produced by K. marxianus CE025. The influence of the temperature in the kinetic parameters was evaluated and best results of ethanol production (12.36 ± 0.06 g.L-1) was achieved at 30 ºC, which is also the optimum temperature for the formation of biomass and the ethanol with a volumetric production rate of 0.25 ± 0.01 g.L-1.h-1 and an ethanol yield of 0.42 ± 0.01 g/g glucose. The results of this study point out the potential of the cashew apple bagasse hydrolysate as a new source of sugars to produce ethanol by S. cerevisiae and K. marxianus CE025. With these results, conclude that the use of cashew apple juice and cashew apple bagasse as substrate for ethanol production will bring economic benefits to the process, because it is a low cost substrate and also solve a disposal problem, adding value to the chain and cashew nut production

Quantificação da floculação de Saccharomyces cerevisiae por bactérias contaminantes da fermentação alcoólica

O assentamento de células de leveduras no fundo das dornas e perdas de células nas centrífugas podem ser causadas por bactérias floculantes, contaminantes naturais da fermentação alcoólica industrial. Estes problemas levam a queda no rendimento e produtividade do etanol. O presente trabalho visa a caracterização da floculação de Saccharomyces cerevisiae por Lactobacillus fermentum CCT 1396. As células de leveduras e bactérias foram misturadas e a floculação das células quantificadas por espectrofotometria. Concentrações de bactérias numa faixa de 0,4 a 3,8g/L (biomassa seca) foram testadas a fim de determinar a ótima concentração de bactérias necessária para provocar a floculação das leveduras. O efeito de pH na floculação das células de leveduras e bactérias foi determinado. 1,38g/L de bactéria foi necessário para a floculação, de 65,4g/L de células de levedura com tempo de contato entre as células (sob agitação) de 15 minutos e repouso de 20 minutos. No pH 3,0 pouco efeito na floculação celular foi detectado e as células continuaram floculadas, mas na faixa de pH 2,0 -- 2,5 a floculação foi próxima de zero. Esta técnica pode ser utilizada para o controle da floculação de leveduras de indústrias de produção de álcool, para determinar a origem desta floculação, já que trata-se de uma técnica fácil, econômica e rápida.

Parâmetros de fermentação ruminal em bovinos alimentados com grãos de milho ou sorgo de alta umidade ensilados

Os parâmetros de fermentação ruminal de dietas contendo silagem de sorgo úmido em substituição à de milho úmido foram estudados em 12 fêmeas bovinas, com peso médio de 584 kg. O delineamento foi inteiramente casualizado com três tratamentos: substituição do milho úmido pelo sorgo úmido ensilado, nos níveis de 0, 50 e 100%. As dietas continham grão úmido de milho ou de sorgo ensilados, soja extrusada, uréia, feno de aveia (Avena sativa sp.), suplemento mineral e monensina. Adicionalmente, foi avaliada a degradabilidade in situ da matéria seca e da fibra em detergente neutro do feno de aveia. Não houve diferença sobre produção total de ácidos graxos voláteis (AGVs) no rúmen, porcentagem molar dos ácidos acético, propiônico e butírico, relação acético/propiônico, pH ruminal, concentração de N-NH3 no rúmen, fluxo e volume de líquidos do rúmen, nos diferentes tratamentos. A degradabilidade da matéria seca e da fibra em detergente neutro do feno não apresentou diferenças. Não se constatou melhora nos parâmetros de fermentação ruminal com a associação dos grãos.

Efeitos de parâmetros de fermentação na produção de etanol a partir de resíduo líquido da industrialização da mandioca (manipueira)

Effect of fermentation parameters on ethanol production from cassava liquid residue (manipueira). "Manipueira", a liquid residue from the processing of cassava starch and flour, has recognized high pollution potential. Aiming at a possible use of "manipueira" as raw material for ethanol production, this study aimed to evaluate the effect of the percentage of inoculated yeast and fermentation temperature on the chromatographic profile of the components of wine of "manipueira". The residue was characterized for starch and sugar content, as well as pH and total acidity. The residue was hydrolyzed by the action of enzymes Termamyl 120 L and AMG 300 L. The hydrolysate obtained was fermented under different experimental conditions. A factorial central composite design (2(2)) with two independent variables and the response surface methodology were used to evaluate the results. The results showed a significant effect of variable parameters on the components of wine. The conditions of low fermentation temperature and lower percentages of inoculated yeast were the most appropriate to obtain ethanol from "manipueira".

Estudo da produção de biossurfactante em caldo de fermentação

A bacterium isolated from soil contaminated by hydrocarbon was studied and, by biochemical tests and analysis of PCR, the presence of Bacillus pumilus was identified. The production of biosurfactant was optimized, test of oil degradation and antimicrobial activity determination. The results showed that pH 5.0 and 7.0, 72 h of fermentation, sucrose and sugar cane juice (2%) had best yields. The bacterium is able to degrade crude oil and displays bacteriostatic and fungistatic activity. From the analysis of proximate composition of biosurfactant found the presence of biopolymer formed by a lipopolysaccharide-protein complex.

Floculação de leveduras por Lactobacillus fermentum em processos industriais de fermentação alcoólica avaliada por técnica fotométrica

Na produção de álcool por fermentação com leveduras, a floculação manifesta-se como um mecanismo natural de agregação de células. Essa condição pode ser induzida por vários fatores, entre eles, a interação entre bactérias floculentas como Lactobacillus fermentum e as leveduras. Esse fato torna-se prejudicial para o processo, pois tanto nas unidades que se utilizam do sistema de recuperação de células por centrifugação, como nas que não o utilizam, ocorrem perdas excessivas de fermento, em conseqüência dos problemas operacionais decorrentes da floculação, comprometendo seriamente o desempenho industrial. No presente trabalho, avaliou-se a floculação em vinho proveniente de fermentação experimental, sob duas condições de pH de fermento tratado utilizadas como inóculo e temperaturas de fermentação, não se observando diferenças significativas entre elas. Com este estudo, pôde-se ainda avaliar a capacidade de dispersão dos flocos em três condições de pH no tratamento do fermento em fase industrial, mostrando diferenças altamente significativas entre elas. A maior capacidade de dispersão no tratamento do fermento é desejável para o controle da floculação industrial, permitindo a sua centrifugação, com conseqüente separação das bactérias contaminantes do fermento. Essas constatações tornaram-se possíveis pelo emprego da técnica usual de determinação da floculação por espectrofotometria, modificada pela desfloculação prévia das amostras de vinho e fermento, o que conferiu maior estabilidade nas leituras.

Padrão de fermentação e composição química de silagens de grãos úmidos de milho e sorgo submetidas ou não a inoculação microbiana

Objetivou-se avaliar o padrão de fermentação e a composição química de silagens de grãos úmidos de milho e de sorgo confeccionadas com ou sem o uso de inoculante microbiano. Avaliou-se, no experimento 1, a silagem de grãos úmidos de milho e, no experimento 2, a silagem de grãos úmidos de sorgo. O material foi ensilado em silos experimentais de PVC (50 cm de comprimento e 100 mm de diâmetro), três por tratamento (tempo de armazenagem, com ou sem inoculante) para cada grão. Amostras foram tomadas antes (0) e aos 1, 2, 3, 4, 8, 16, 32 e 64 dias após a ensilagem, totalizando 48 silos experimentais para cada grão. Os tratamentos foram arranjados em delineamento inteiramente casualizado, em esquema fatorial 2 x 9 (com e sem inoculante microbiano, nove tempos de armazenagem), com três repetições para cada grão. Não houve efeito da inoculação e do tempo pós-ensilagem sobre o teor de MS dos grãos úmidos de milho e de sorgo, com médias de 64,13 e 64,03% e de 67,66 e 67,48% para silagens controle e inoculadas, respectivamente. Não houve efeito da inoculação sobre o pH dos grãos úmidos de milho e de sorgo aos 64 dias após ensilagem, com médias de 3,97 e 3,92 e de 3,94 e 3,95 unidades para silagens controle e inoculadas, respectivamente. O inoculante microbiano não promoveu alterações na composição química nem redução de perdas da MS nas silagens de grãos úmidos de milho e de sorgo. Nas condições estudadas, não é necessária inoculação para melhoria nos padrões fermentativos de silagens de grãos úmidos de milho e de sorgo.

Influência da utilização de aditivos químicos no perfil da fermentação, no valor nutritivo e nas perdas de silagens de cana-de-açúcar

Avaliou-se o efeito do uso de aditivos químicos nas perdas de matéria seca, no perfil fermentativo e no valor nutritivo de silagens de cana-de-açúcar. O experimento foi realizado em delineamento inteiramente casualizado com quatro repetições por tratamento (sete tratamentos). Os seguintes aditivos foram utilizados na confecção das silagens: controle (sem aditivo), L. buchneri, óxido e carbonato de cálcio em doses de 1,0 e 1,5% da massa verde e sulfato de cálcio a 1,0% da massa verde, diluídos em 40 litros de água por tonelada de forragem. As variáveis analisadas foram: perdas totais e gasosas, produção de efluente, recuperação de matéria seca, composição química e valor nutritivo. As menores perdas fermentativas e gasosas foram observadas nas silagens com óxido ou carbonato de cálcio, que resultaram em maior taxa de recuperação de matéria seca. da mesma forma, as silagens tratadas com estes aditivos apresentaram maior teor de carboidratos solúveis residuais e de ácido lático e reduzida fermentação alcoólica. As silagens tratadas com óxido e carbonato apresentaram, no momento da abertura, maior teor de cinzas, menor concentração de componentes fibrosos e maiores coeficientes digestibilidade da matéria seca e da matéria orgânica. O teor de proteína encontrado nessas silagens foi semelhante ao observado na forragem fresca. A ensilagem de cana-de-açúcar com L. buchneri e sulfato de cálcio possibilitou a obtenção de desempenho similar ao determinado com a silagem sem aditivos. Silagens tratadas com carbonato e óxido de cálcio apresentam maior desempenho durante o processo fermentativo.

Zymomonas mobilis: um microrganismo promissor para a fermentação alcoólica

In many countries, fermentation studies regarding the use of bacteria instead of yeasts to reduce the period of alcoholic fermentation have been carried out. In Brazil, all the industrial alcohol production is carried out by yeasts as fermentation microorganisms and little is known about other microorganisms with potential to produce alcohol industrially. Brazil stands out in the energy sector worldwide and thus some institutions have been selecting microorganisms which are more efficient in the alcohol production process. Alcoholic bacteria from species Zymomonas mobilis present technological characteristics with potential to be used for alcoholic fermentation at industrial scale, since it exhibits promising abilities to transform sugars into alcohol and carbon dioxide, at conditions similar to the ones required by yeasts. Zymomonas mobilis is a unique bacteria among the microbial world, with peculiar growth, energy production and response to culture conditions, causing a great interest in scientific, biotechnological and industrial fields. The bacteria's ability to make possible energy production in favor of product formation, respond to physical and chemical environmental manipulation as well as its limited product formation make it an ideal microorganism for the study and development of microbial processes for ethanol production.