Effects of feeding corn silage inoculated with microbial additives on the ruminal fermentation, microbial protein yield, and growth performance of lambs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ethanol production by Zymomonas mobilis during sucrose fermentation: Optimization of culture conditions using factorial design

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purification and properties of polygalacturonase produced by thermophilic fungus Thermoascus aurantiacus CBMAI-756 on solid-state fermentation

Polygalacturonases are enzymes involved in the degradation of pectic substances, being extensively used in food industries, textile processing, degumming of plant rough fibres, and treatment of pectic wastewaters. Polygalacturonase (PG) production by thermophilic fungus Thermoascus aurantiacus on solid-state fermentation was carried out in culture media containing sugar cane bagasse and orange bagasse in proportions of 30% and 70% (w/w) at 45°C for 4 days. PG obtained was purified by gel filtration and ion-exchange chromatography. The highest activity was found between pH 4.5 and 5.5, and the enzyme preserved more than 80% of its activity at pH values between 5.0 and 6.5. At pH values between 3.0 and 4.5, PG retained about 73% of the original activity, whereas at pH 10.0 it remained around 44%. The optimum temperature was 60–65°C. The enzyme was completely stable when incubated for 1 hour at 50°C. At 55°C and 60°C, the activity decreased 55% and 90%, respectively. The apparent molecular weight was 29.3 kDa, Km of 1.58 mg/mL and Vmax of 1553.1μmol/min/mg. The presence of Zn+2, Mn+2, and Hg+2 inhibited 59%, 77%, and 100% of enzyme activity, respectively. The hydrolysis product suggests that polygalacturonase was shown to be an endo/exoenzyme.

Bioconversion of Cellulose to Hydrogen by dark fermentation

Hydrogen is known as a clean energy resource. The biological production of hydrogen has been attracting attention as an environmentally friendly processs that does not consume fossil fuels. Cellulosic plant and waste materials are potential resources for fermentative hydrogen production. Cellulose is a linear biopolymer of glucose molecules, connected by β-1,4-glycosidic bonds. Enzymatic hydrolysis of cellulose requires the presence of cellulase. The present study aimed to investigate the efficiency of acid pretreatment on ruminal fluid in order to enrich H2 producing bacteria consortia to enhance biohydrogen rate and substrate removal efficiency. In this study, fermentative hydrogen producers were enriched on cellulose (2g/L) in a modificated Del Nery medium (DNM) at 37ºC and initial pH 7.0 using rumen fluid (10% v/v) as inoculum. To increase the hydrogen production it was added cellulose (10mL) to the medium. The gas products (mainly H2 and CO2) was analyzed by gas chromatography (Shimadzu GC 2010) using a thermal conductivity detector. The volatile fatty acids and ethanol were also detected by GC using a flame ionization detector. Cellulose degradation was quantified by using the phenolsulfuric acid method. Analysis showed that the biogas produced from the anaerobic fermentation contained only hydrogen and carbon dioxide, without detectable methane after acid pretreatment test. On DNM the hydrogen production started with 4 h (5,3 x 105 mmol H2/L) of incubation, and the maximum H2 concentration was observed with 34 h (7,1 x 106 mmol H2/L) of incubation. During the process, it was observed a predominance of acetic acid and butyric acid as well as a low production of acetone, ethanol and nbutanol in all experimental phases. Butyrate accounted for more than 77% of total. As a result of the accumulation of volatile fatty acids (VFAs), the pH value in anaerobic digestion system was reduced to 4,0. On microscopy analyses there were observed rods with endospores. The batch anaerobic fermentation assays performed on anaerobic mixed inoculum from rumen fluid demonstrated the feasibility of H2 generation utilizing cellulose as substrate. Based on the results, it can be concluded that the acid treatment was efficient to inhibit the methanogenic archaea cells present in rumen fluid. The rumen fluid cells present a potential route in converting renewable biomass such as cellulose into hydrogen energy.

Estimation of Submerged Aquatic Vegetation Height and Distribution in Nova Avanhandava Reservoir (São Paulo State, Brazil) Using Bio-Optical Modeling

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Green and brown propolis: efficient natural biocides for the control of bacterial contamination of alcoholic fermentation of distilled beverage

This study aimed to evaluate the efficiency of natural biocides, brown and green propolis, for the control of bacterial contamination in the production of sugarcane spirit. The treatments consisted of brown and green propolis extracts, ampicillin, and a control and were assessed at the beginning and end of harvest season in ten fermentation cycles. In the microbiological analyses, the lactic acid bacteria were quantified in the inoculum before and after the treatment with biocides, and the viability of yeast cells during fermentation was evaluated. The levels of acids, glycerol, total residual reducing sugars, and ethanol were analyzed for the wine resulting from each fermentation cycle. A reduction in the number of bacterial contaminants in the inoculum in the treatments with the natural biocides was observed, but it did not affect the viability of yeast cells. The control of the contaminants led to the production of higher levels of ethanol and reduced acidity in the wine produced. The results of the use of brown and green propolis to control the growth microorganisms in the fermentation of sugarcane spirit can be of great importance for using alternative strategies to synthetic antibacterials in fermentation processes including other distilled beverage or spirits.

Marine-derived fungi: diversity of enzymes and biotechnological applications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hygroscopic properties of solid agro-industrial by-products used in solid-state fermentation

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mathematical modeling of a continuous alcoholic fermentation process in a two-stage tower reactor cascade with flocculating yeast recycle

Experiments of continuous alcoholic fermentation of sugarcane juice with flocculating yeast recycle were conducted in a system of two 0.22-L tower bioreactors in series, operated at a range of dilution rates (D (1) = D (2) = 0.27-0.95 h(-1)), constant recycle ratio (alpha = F (R) /F = 4.0) and a sugar concentration in the feed stream (S (0)) around 150 g/L. The data obtained in these experimental conditions were used to adjust the parameters of a mathematical model previously developed for the single-stage process. This model considers each of the tower bioreactors as a perfectly mixed continuous reactor and the kinetics of cell growth and product formation takes into account the limitation by substrate and the inhibition by ethanol and biomass, as well as the substrate consumption for cellular maintenance. The model predictions agreed satisfactorily with the measurements taken in both stages of the cascade. The major differences with respect to the kinetic parameters previously estimated for a single-stage system were observed for the maximum specific growth rate, for the inhibition constants of cell growth and for the specific rate of substrate consumption for cell maintenance. Mathematical models were validated and used to simulate alternative operating conditions as well as to analyze the performance of the two-stage process against that of the single-stage process.

Effect of the particle size of maize, rice, and sorghum in extruded diets for dogs on starch gelatinization, digestibility, and the fecal concentration of fermentation products

The influence of rice, maize, and sorghum raw material particle size in extruded dry dog food on the digestibility of nutrients and energy and the fecal concentration of fermentation products was investigated. Three diets with similar nutrient compositions were formulated, each with 1 starch source. Before incorporation into diets, the cereals were ground into 3 different particle sizes (approximately 300, 450, and 600 mu m); therefore, a total of 9 diets were in a 3 x 3 factorial arrangement (3 cereals and 3 particle sizes). Fifty-four beagle dogs (12.0 +/- 0.1 kg BW) were randomly assigned to the diets, with 6 dogs per diet. The digestibility was measured with the chromium oxide method. The data were evaluated with ANOVA considering the carbohydrate source, grinding effect, and interactions. The means were compared with the Tukey test and polynomial contrasts (P < 0.05). With the same grinding procedure, rice was reduced to smaller particles than other cereals. The cereal mean geometric diameter (MGD) was directly related to starch gelatini-zation (SG) during extrusion. For rice diets, the MGD and SG did not change nutrient digestibility (P > 0.05); only GE digestibility was reduced at the largest MGD (P < 0.01). For maize and sorghum diets, the total tract apparent nutrient digestibility was reduced for foods with greater MGD and less SG (P < 0.01). A linear reduction in nutrient digestibility according to cereal particle size was observed for sorghum (r(2) < 0.72; P < 0.01). Higher concentrations of fecal total shortchain fatty acids (SCFA) were observed for sorghum diets (P < 0.05) than for other diets. The rice diets led to the production of feces with less lactate (P < 0.05). The increase in raw material MGD did not influence fecal SCFA for rice diets, but for the dogs fed maize and sorghum foods, an increase in propionate and butyrate concentrations were observed as MGD increased (P < 0.05). In conclusion, for dogs fed different particle sizes of the cereal starches in the extruded diets, the digestibility and fecal characteristics were affected, and this effect was ingredient dependent.

Feeding increasing concentrate to Tifton 85 hay ratios modulated rumen fermentation and microbiota in Nellore feedlot steers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Improvement of submerged culture conditions to produce colorants by Penicillium purpurogenum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of supplementation frequency on the ruminal fermentation and enteric methane production of beef cattle grazing in tropical pastures

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of grass carp (ctenopharyngodon idella) as a biological control agent for submerged aquatic macrophytes

This study aimed to evaluate feed preference and control efficacy of grass carp (Ctenopharyngodon idella) on the aquatic macrophytes Ceratophyllum demersum, Egeria densa and Egeria najas. An experiment was carried out at mesocosms conditions with 2,000 liters capacity and water residence time of 2.8 days. C. demersum, E. densa e E. najas biomasses were offered individually with sixty g and coupled in similar quantities of 30 g of each species, evaluated during 81 days, envolving 6 treatments. (1 - C. demersum, 2 - E. najas, 3 - E. densa, 4 - C. demersum + E. najas, 5 - C. demersum + E. densa and 6 - E. najas + E. densa). When offered individually, E. najas and C. demersum presented the same predation rate by grass carp, which was higher than E. densa predation rate. When plants were tested in pairs, the order of feed preference was C. demersum > E. najas > E. densa. E. najas and C. demersum percentage control ranged from 73 to 83%. No relation between biomass consumption and grass carp body weight gain was observed, probably due to differences in nutritional quality among macrophyte species according to fish necessities. Therefore, it is concluded that the use of grass carp is one excellent technique to control submersed macrophytes in Brazil.

Purification and characterization of a unique pectin lyase from aspergillus giganteus able to release unsaturated monogalacturonate during pectin degradation

A pectin lyase, named PLIII, was purified to homogeneity from the culture filtrate of Aspergillus giganteus grown in submerged culture containing orange peel waste as carbon source. PLIII was able to digest apple pectin and citrus pectins with different degrees of methyl esterification. Interestingly, the PLIII activity was stimulated in the presence of some divalent cations including Pb(2+) and was not significantly affected by Hg(2+). Like other pectin lyases, PLIII is stimulated by but is not dependent on Ca(2+). The main soluble product released during the degradation of pectic substances promoted by the PLIII is compatible with an unsaturated monogalacturonate. PLIII is a unique enzyme able to release unsaturated monogalacturonate as the only soluble product during the degradation of pectic substances; therefore, PLIII was classified as an exo-pectin lyase. To our knowledge, this is the first characterization of an exo-pectin lyase. The PLIII described in this work is potentially useful for ethanol production from pectin-rich biomass, besides other common applications for alkaline pectinases like preparation of textile fibers, coffee and tea fermentation, vegetable oil extraction, and the treatment of pulp in papermaking.