Effect of Partial and Total Replacement of Inorganic by Organic Microminerals Sources on the Quality of Broiler Carcasses

ABSTRACT Among the nutrients involved in the chicken diet, the microminerals deserve attention as they exert essential functions in the organism. These compounds can be provided in inorganic (traditional) and organic (chelate) forms. In organic form, the micromimerals can attend a new concept related to a better bio-availability. In this sense, the objective of this work was to evaluate the effect of the partial and total substitution of inorganic by organic sources of microminerals on the animal diet, assessing its concentration in the blood and liver after 21 and 40 days and also in the bones after 40 days. Moreover, the effect on the physical-chemical and sensory characteristics of the muscle after 15 days of storage at 4°C was also evaluated. The animals were divided into five treatments: 100% inorganic, 70% inorganic and 30% organic, 50% inorganic and 50% organic, 30% inorganic and 70% organic and 100% organic-chelate. It is shown that the use of organic sources of microminerals in the diet of poultry induced to an increase of its concentrations in the blood and liver, with no significant alterations in the compositions of bones, compared to the use of inorganic sources. Feed formulated using 50% of organic minerals and 50% of inorganic minerals led to similar results. The diet with organic minerals or mixture with inorganic sources results in a low lipid oxidation in the drumsticks stored at 4°C for 15 days, in comparison with those using only inorganic minerals. No sensory alterations were observed for all different treatments.

Characterization of novel thermostable polygalacturonases from Penicillium brasilianum and Aspergillus niger.

The aim of this research was the partial characterization of polygalacturonase (PG) extracts produced by a newly isolated Penicillium brasilianum and Aspergillus niger in submerged fermentation. The partial characterization of the crude enzymatic extracts showed optimum activity at pH 5.5 and 37 °C for both extracts. The results of temperature stability showed that PG from both microorganisms were more stable at 55 °C. However, the enzyme obtained by P. brasilianum presents a half-life time (t 1/2 = 693.10 h), about one order of magnitude higher than those observed in for A. niger at 55 °C. In terms of pH stability, the PG produced by P. brasilianum presented higher stability at pH 4.0 and 5.0, while the PG from A. niger showed higher stability at pH 5.0.

Elaboração e avaliação da qualidade de macarrão isento de glúten / Elaboration and quality evaluation of the gluten-free pasta

Com a intenção de minimizar a carência de produtos isentos de glúten e tendo em vista o crescente aumento de celíacos, a elaboração de produtos diferenciais a base de farinha de arroz e milho torna-se uma alternativa de consumo e comercialização. Assim, este trabalho teve como objetivo avaliar os efeitos da adição de farinha de arroz e milho, e emulsificante nas características de qualidade (teste de cozimento, textura instrumental - firmeza e aceitabilidade) e físico-químicas (proteínas, lipídios, carboidratos, minerais e fibra alimentar) de formulações de macarrão, mediante o emprego de metodologia de planejamento de experimentos, fixando a concentração de ovos in natura (12%, m/m) e água (20%, v/m). A formulação com 40g de farinha de milho, 60g de farinha de arroz e 0,4g de SSL por 100g de farinha mista foi a que apresentou melhor resistência ao cozimento, menores perdas de sólidos na água de cocção (2,12% m/m), textura (30,40g seg-1) e aceitabilidade superior a 85 %. Além disso, o uso de farinha de arroz, associado à farinha de milho, proporciona fonte de minerais (152mg de K, 1,55mg de Fe, 46mg de Mg, 20mg de Ca por 100g), proteínas (~7%), carboidratos (52%) e fibras (~1%), importante suprimento nutricional para celíacos, por estes apresentarem perdas significativas destes nutrientes, devido à diarreia e à absorção restrita, característica da doença.

In order to minimize the deficiency of products with gluten-free and the growing number of celiac the elaboration of differential products using rice and maize flour becomes an alternative of consuming and marketing. This study aimed to evaluate the effects of rice and maize flour, and emulsifier addition in the quality characteristics (cooking test, instrumental texture - firmness and acceptability) and physic-chemical (proteins, lipids, carbohydrates, minerals and dietary fiber) of pasta formulation, by the use of experimental design methodology, fixing the concentration of in nature eggs (12%, w/w) and water (20%, v/w). The formulation with 40g of maize flour, 60g of rice flour and 0.4g per 100g of SSL mixed flour showed the better cooking resistance, reduced loss of solids in the cooking water (2.12% m/m), texture (30.40g sec-1) and acceptability higher than 85%. Furthermore, the use of rice flour associated with maize flour provide a source of minerals (152mg of K, 1.55mg of Fe, 46mg of Mg, 20mg Ca per 100g), proteins (~ 7%), carbohydrates (52%), fibers (~1%) important nutritional supplement for celiac, because they present significant losses of these nutrients due to restricted absortion and diarrhea, characteristic of the disease.

Synthesis of a hybrid polymer-inorganic biomimetic support incorporating in situ pectinase from Aspergillus niger ATCC 9642.

The hybrid alginate/gelatin/calcium oxalate (AGOCa) support was successfully synthesized through the biomimetic mineralization method for immobilization in situ of a pectinolytic extract from Aspergillus niger ATCC 9642 via entrapment technique. The efficiency of immobilization reached 72.7%. Sodium oxalate buffer (100 mM, pH 5.5) was selected as adjuvant of the immobilization process by allowing the formation of a calcified shell around the calcium alginate capsule, significantly increasing the stability to storage, thermal and recycling of the enzymatic immobilized pectinolytic extract. The pH and temperature for maximum activity were from 5.0 to 6.0 and 60 to 80 °C, respectively. The new hybrid support can be a potential alternative to obtain immobilized pectinases with properties for advantageous industrial applications.

Control of Penicillium sp. on the Surface of Italian Salami Using Essential Oils.

The goal of this study is to evaluate the in vitro effects of rosemary, salvia, oregano and clove oils at volume fractions of 1000, 750, 500, 250, 100, 50, 26, 10 and 5 µL/mL (100, 75, 50, 25, 10, 5, 3, 1 and 0.5%) on the growth of contaminating fungi in salami. The in vitro effect of the oils against fungal growth was indicated by zones of inhibition. Rosemary oil showed an inhibition zone of 9.6 mm only at the maximal volume fraction (1000 µL/mL). Salvia oil showed inhibition zones of 12.2, 11.2 and 10.5 mm only at the three highest fractions tested. Based on the inhibition zones, clove oil at 125 and 250 µL/mL, oregano oil at 250 and 500 µL/mL and a mixture (1:1 by volume) of the two oils at 100 µL/mL were selected to be applied to the surface of salamis. A significant reduction of fungal growth in all of the oil-treated samples was confirmed by visual inspection. A sensory analysis revealed that the samples treated with 125 µL/mL of clove oil or 100 µL/mL of a mixture of oregano and clove oil showed no significant flavour differences compared with the control. Carvacrol and eugenol were the principal compounds in oregano and clove oils, respectively, and were most likely responsible for the antifungal activity.

Decontamination of Pig Carcasses Using Water Pressure and Lactic Acid

The objective of this work was to evaluate the effect of different water pressures and concentrations of lactic acid on microbial counts (mesophilic bacteria, enterobacteria and Salmonella) on pig carcasses without contamination and contaminated carcasses, before and after the last shower and before being cooled. The tests were carried out using 4, 3 and 2 bar water pressure, and 2, 1 and 0% lactic acid concentration. In general, both the pressure in the shower and lactic acid had a positive effect by reducing the microbial count. The interaction between the pressure and lactic acid caused the largest reduction in carcasses surface count for mesophiles. With regard to enterobacteria on contaminated carcasses, the most important variable was the lactic acid concentration and in uncontaminated carcasses, it was water pressure. The use of 8 bar pressure of the wash water without lactic acid caused a reduction in mesophilic bacteria and enterobacteria, for both the contaminated and uncontaminated carcasses, with results statistically equal to each other, and significantly lower than the initial counts. The water pressure at 8 bar reduced the percentage of carcasses with Salmonella in contaminated carcasses.

Experimental Design Applied to the Optimization and Partial Characterization of Pectin Liase from a Newly Isolated Penicillium brasilianum

Penicillium brasilianum was previously isolated from tea and identified by molecular biology technique. A Plackett-Burman design, followed by a complete second order design was used for the screening of most important factors and to maximize the pectin liase (PMGL) activity, respectively. The maximum PMGL activity by P. brasilianum achieved was 9.0 U/mL after 48 h of cultivation in a medium containing pectin (33.0 g/L), yeast extract (30.0 g/L) and potassium phosphate (2.0 g/L) at 30ºC, with a stirring rate of 180 rpm, initial pH 5.5 and 5x106 spores/mL inoculum size. The kinetic evaluation in terms of substrate consumption demonstrated that the maximum production of PMGL was at 72 h, and 40% of the total organic carbon, 25% of the nitrogen, 88% of the magnesium, 13% of the potassium and 66% of the iron were consumed. The pH remained almost stable during the whole period of production (5.33 to 4.9). The partial characterization of the crude PMGL enzyme extract showed optimal pH and temperature of 5.5 and 37°C, respectively.

Comparative lethality kinetic curves and predictive models of F-value for Listeria monocytogenes using different sanitizers.

The objective of this work was to evaluate the kinetic of inactivation of Listeria monocytogenes using peracetic acid, chlorhexidine, and organic acids as active agent, determining the respective D-, Z-, and F-values. From our knowledge, these important results from an industrial view point are not available in the current literature, mainly for organic acids, pointing out the main contribution of the present work. Lower D-values were obtained for peracetic acid and chlorhexidine, compared with the organic acids. For the reduction of 6 log10 of L. monocytogenes using peracetic acid, at 0.2, 0.1, and 0.05% are necessary 7.08, 31.08, and 130.44 min of contact, respectively. The mathematical models of F-values showed that at concentrations lower than 0.15% one can verify an exponential increase in F-values, for both de chlorhexidine and peracetic acid. The organic acids presented a linear behavior, showing slight variation in F-values, is even more effective in under dosage. The results obtained are of fundamental importance in terms of industrial strategy for sanitization procedure, permitting to choose the best relation product concentration/exposure time, aiming at reducing costs without compromising the disinfectant efficiency.

Ultrasound-assisted enzymatic transesterification of methyl benzoate and glycerol to 1-glyceryl benzoate in organic solvent.

The aim of this work is to report the enzymatic transesterification production of 1-glyceryl benzoate under ultrasound irradiation, using a commercial immobilized lipase, Novozym 435. Firstly, a preliminary evaluation was carried out at 2, 4 and 6h, at constant temperature of 50 °C, methyl benzoate to glycerol molar ratio of 1:1 and 5.5 wt% of enzyme concentration. After analyzing the results obtained, the experimental design technique was used to evaluate the effects of temperature, substrates molar ratio, enzyme concentration, solvent volume and ultrasonic power on the 1-glyceryl benzoate production. The highest conversion, around 16%, was obtained at 65 °C, 1:1 of methyl benzoate to glycerol molar ratio, 15 wt% of enzyme concentration, 7 mL of solvent and 40% ultrasonic power in 4h of reaction. A preliminary kinetic experiment carried out varying the enzyme concentration (15 and 20 wt%) keeping fixed the temperature at 35 °C, 1:1 of substrates molar ratio, 3 mL of solvent and 40% of maximum ultrasonic power led to lower (around 15% after 12 h of reaction) conversions compared to that achieved in the experimental design.

Enzymatic synthesis of ascorbyl palmitate in ultrasound-assisted system: process optimization and kinetic evaluation.

This work is focused on the optimization of reaction parameters for the synthesis of ascorbyl palmitate catalyzed by Candida antarctica lipase in different organic solvents under ultrasound irradiation. The sequential strategy of experimental design proved to be useful in determining the optimal conditions for reaction conversion in tert-butanol system using Novozym 435 as catalyst. The optimum production was achieved at 70°C, ascorbic acid to palmitic acid molar ratio of 1:9, enzyme concentration of 5 wt% at 3h of reaction, resulting in an ascorbyl palmitate conversion of about 27%. Reaction kinetics for ascorbyl palmitate production in ultrasound device showed that satisfactory reaction conversions (∼26%) could be achieved in short reaction times (2h). The empirical kinetic model proposed is able to satisfactorily represent and predict the experimental data.

Optimization of α-Terpineol production by the biotransformation of R-(+)-limonene and (-)-ß-pinene.

The main objective of this work is to present the optimization of the biotransformation of R-(+)-limonene and (-)-ß-pinene aiming at the production of α-terpineol by strains of fungal and yeasts previously isolated by our research group using the methodology of experimental design. New optimized experimental data on α-terpineol production by the biotransformation of R-(+)-limonene and (-)-ß-pinene using newly isolated microorganisms are reported in this work. Conversion of about 1,700 mg/L was achieved when R-(+)-limonene was used as substrate and the newly isolated strain 05.01.35 as microorganism at the central point of the experimental design, corresponding to a substrate concentration of 1.75%, mass of inoculum of 2 g, and substrate to ethanol volume ratio of 1:1. The same experimental condition led to higher conversions when (-)-ß-pinene was used as substrates and the strains coded as 04.05.08 and 01.04.03 as microorganism. Here, conversions of about 770 mg/L were achieved.

Insecticidal and repellency activity of essential oil of Eucalyptus sp. against Sitophilus zeamais Motschulsky (Coleoptera, Curculionidae).

BACKGROUND: The objective of this work was to evaluate the insecticidal and repellent effect of essential oils of Eucalyptus dunnii, E. saligna, E. benthamii, E. globulus and E. viminalis in the control of Sitophilus zeamais in maize grains. RESULTS: The essential oils chemical composition showed that the E. benthamii presents the cis-ocimene (56.88%) as major compound. Results permitted us to verify that the evaluated species presented different insecticidal action under Sitophilus zeamais. About 100% of mortality was achieved with doses of 65, 100 and 400 microL for E. dunnii, E. saligna and E. benthamii, respectively. After regression analysis the LD(50) values were calculated and E. dunnii was shown to be the most efficient (25.03 microL), followed by E. saligna (37.93 microL) and E. benthamii (121.09 microL). Using the previously calculated LD(50) values, the repellency activity was calculated, allowing us to conclude that all species presented significant values in terms of this important parameter. CONCLUSION: The essential oil of Eucalyptus sp. presented insecticidal and repellency against Sitophilus zeamais Motschulsky (Coleoptera, Curculionidae). Although from an economic point of view synthetic chemicals are still more frequently used as repellents, natural products (essential oils) have the potential to provide efficient and safer repellents for humans and the environment.

Screening of pectinase-producing microorganisms with polygalacturonase activity.

The aim of this work was to perform the screening of microorganisms, previously isolated from samples of agro-industrial waste and belonging to the culture collection of our laboratory, able to produce polygalacturonases (PG). A total of 107 microorganisms, 92 newly isolated and 15 pre-identified, were selected as potential producers of enzymes with PG activity. From these microorganisms, 20 strains were able to synthesize PG with activities above 3 U mL(-1). After the kinetic study, the enzyme activity was increased up to 13 times and the microorganism identified as Aspergillus niger ATCC 9642 and the newly isolated W23, W43, and D2 (Penicillium sp.) after 24 h of fermentation led to PG activities of 30, 41, 43, and 45 U mL(-1), respectively. The RAPD analysis demonstrated that the selected strains differs genetically, indicating that no duplication of strains among them in the experiments for polygalacturonases production was verified.

Microbial oxidation of (-)-α-pinene to verbenol production by newly isolated strains.

Verbenol is a bicyclicbicycle secondary allylic alcohol, with pronounced camphor and mint flavor notes, mainly used as food flavoring. This compound is also used to control harmful insects, and hence has potential for using in agriculture, and is an intermediate in the synthesis of valuable perfume and medicinal substances. This work is focused on the microbial oxidation of (-)-α-pinene to verbenol production. To carry out the present study, 405 microorganisms were tested for their ability to bioconvert the substrate. From the isolated microorganisms, 193 were selected in the pre-screening using mineral medium for limonene degradation. At the screening step, 31 strains were able to convert (-)-α-pinene in verbenol. The highest concentration in verbenol from (-)-α-pinene was about 125.6 mg/L for yeast isolated from orange juice industrial residue.

Isolation and screening of microorganisms for R-(+)-limonene and (-)-beta-pinene biotransformation.

This work is focused on the biotransformation of R-(+)-limonene and (-)-beta-pinene to bioflavor production. To carry out the present study, 405 microorganisms were tested for their ability to bioconvert the substrates. From the isolated microorganisms, 193 were selected in the prescreening using mineral medium for limonene degradation. At the screening step, eight strains were able to convert R-(+)-limonene and 15 to transform (-)-beta-pinene, both in alpha-terpineol. The highest concentration in alpha-terpineol from R-(+)-limonene was about 3,450 mg/L for Penicillium sp. isolated from eucalyptus steam. From (-)-beta-pinene, the highest product concentration of 675.5 mg/L was achieved using an Aspergillus sp. strain isolated from orange tree stem.

Enzyme-catalyzed production of 1-glyceryl benzoate in compressed n-butane.

The aim of this work is to report the enzymatic transesterification production of 1-glyceryl benzoate in compressed n-butane, using a commercial immobilized lipase, Novozym 435. For this purpose, reaction experiments were performed on the basis of phase equilibrium data of the system methyl benzoate/n-butane, measured using the static synthetic method with a variable-volume view cell in the temperature range of 313.15-343.15K and pressures up to 12MPa, in the entire compositional range of n-butane. Results indicate the existence of a relatively complex phase behavior for all temperatures investigated with the occurrence of vapor-liquid and liquid-liquid phase transitions. Reaction results showed that the strategy adopted for the experimental design proved to be useful in optimizing the reaction conversion in pressurized n-butane and Novozym 435. The optimum conditions were found to be 5.5wt% of enzyme, methyl benzoate to glycerol molar ratio of 3:1, 50°C and 6h of reaction, affording about 6% of 1-glyceryl benzoate yield.

Microorganism screening for limonene bioconversion and correlation with RAPD markers.

The use of microorganisms for biotransformations of monoterpenes has stimulated the biotechnological market. Aiming at the highest efficiency in the process of strains screening, the application of molecular biology techniques have been proposed. Based on these aspects, the objective of this work was to select different strains able to convert limonene using fermentative process and random amplified polymorphic DNA (RAPD) markers. The results obtained in the fermentative screening, from 17 strains tested, pointed out that four microorganisms were able to convert limonene into oxygenated derivatives. The RAPD study showed a polymorphism of 96.02% and a similarity from 16.02 to 51.51%. Based on this it was possible to observe a high genetic diversity, even among strains of same species, concluding that the RAPD was not able to correlate the genetic characteristics of the microorganism with the results obtained from the biotransformation process.

Biotransformation of (-)beta-pinene by Aspergillus niger ATCC 9642.

The main objective of this work was to investigate the biotransformations of (-)alpha-pinene, (-)beta-pinene, and (+) limonene by Aspergillus niger ATCC 9642. The culture conditions involved--concentration of cosolvent (EtOH), substrate applied, and sequential addition of substrates were--investigated. Adaptation of the precultures with small amounts of substrate was also studied. The experiments were performed in conical flasks with liquid cultures. This strain of A. niger was able to convert only (-)beta-pinene into alpha-terpineol. An optimum conversion of (-)beta-pinene into alpha-terpineol of about 4% was obtained when the substrate was applied as a diluted solution in EtOH and sequential addition of substrate was used.