Effect of phytase in laying hen diets with different phosphorus sources

An experiment was conducted to evaluate the effects of the enzyme phytase in diets formulated with different phosphorus sources on performance, eggshell quality and excretion of commercial laying hens. Two hundred and eighty-eight commercial Hyssex Brown laying hens were evaluated during two production phases, which included eight twenty-eight-day cycles, using a completely randomized design in a 3x2 factorial with six replicates of eight birds per treatment. Three phosphorus sources (calcium and sodium phosphate, micro-granulated dicalcium phosphate and triple super phosphate) and two phytase levels (0 or 1000 FTU/kg diet) were tested in the composition of the diets. After the post-peak period, triple super phosphate decreased bird performance and eggshell quality. It was possible to reduce the levels of phosphorus supplementation when phytase was added to the diet. Besides, phytase supplementation reduced phosphorus, calcium and nitrogen excretions, but affected mean egg weight at production peak.

Enzyme use in kibble diets formulated with wheat bran for dogs: Effects on processing and digestibility

Recently, there is an interest in technologies that favour the use of coproducts for animal nutrition. The effect of adding two enzyme mixtures in diets for dogs formulated with wheat bran (WB) was evaluated. Two foods with similar compositions were formulated: negative control (NC; without WB) and test diet (25% of WB). The test diet was divided into four treatments: without enzyme (positive control), enzyme mixture 1 (ENZ1; added before extrusion β-glucanase, xylanase, cellulase, glucoamylase, phytase); enzyme mixture 2 (ENZ2; added before extrusion the ENZ1 more α-amylase); enzyme mixture 2 added after the extrusion (ENZ2ex). ENZ1 and ENZ2 were used to evaluate the enzyme effect on extruder pre-conditioner (processing additive) and ENZ2ex to evaluate the effect of enzyme supplementation for the animal. Digestibility was measured through total collection of faeces and urine. The experiment followed a randomized block design with five treatments (diets) and six dogs per diet, totalling 30 dogs (7.0 ± 1.2 years old and 11.0 ± 2.2 kg of body weight). Data were submitted to analysis of variance and means compared by Tukey's test and orthogonal contrasts (p < 0.05). Reducing sugars showed an important reduction after extrusion, suggesting the formation of carbohydrate complexes. The apparent total tract digestibility (ATTD) of dry matter, organic matter, crude protein, acid-hydrolysed fat and energy was higher in NC than in diets with WB (p < 0.001), without effects of enzyme additions. WB diets resulted in higher faecal production and concentration of short-chain fatty acids (SCFA) and reduced pH and ammonia concentration (p < 0.01), with no effect of enzyme addition. The enzyme addition did not result in improved digestibility of a diet high in non-starch polysaccharides; however, only ATTD was measured and nutrient fermentation in the large intestine may have interfered with the results obtained. WB modified fermentation product formation in the colon of dogs. © 2013 Blackwell Verlag GmbH.

Effect of phytase supplementation on performance, bone densitometry and carcass yield in broilers chicks

An experiment was conducted to evaluate the performance, bone densitometry and carcass yield of broilers chicks, using different levels of phytase enzyme. Nine hundred and sixty male one-day-old broiler chicks were used. The birds were distributed in a completely randomized experimental design, involving five treatments and six replications of 32 chicks each. The treatments consisted of a control diet for each phase, and four other diets were formulated adding growing levels of the phytase enzyme (250, 500, 750 and 1,000 FTU of phytase kg-1 feed). When adding the phytase enzyme, the nutritional matrix was valued to guarantee the same nutritional levels as the control diet. In general, the addition of phytase enzyme determined a linear decrease on the performance of the birds. However, the performance obtained with the level of 250 FTU phytase kg-1 feed were no different from the control treatment. The best bone density results were observed in the control treatment with no phytase, and the highest leg and thigh yield were obtained at the level of 514 FTU phytase kg-1.

Phytase production by Rhizopus microsporus var. microsporus biofilm: characterization of enzymatic activity after spray drying in presence of carbohydrates and nonconventional adjuvants

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

Biotechnology in the development of improved phytases

Phytase enzyme supplements are now ubiquitous in the commercial production of a range of livestock, particularly chickens and pigs. Significant effort has been directed over the last two decades towards producing improved enzymes with higher activity, increased stability and at economic levels in industrial fermentations. As such, there are excellent products on the market, but there is a continuing demand for further improvements to drive down costs and for enzyme manufacturers to increase market share. The rapid development of DNA sequencing and gene synthesis technologies has provided ready access to a large number of new and uncharacterised potential phytases. Challenges remain however in identifying and developing those with improved properties.

Enzimas exógenas em dietas de frangos de corte: desempenho

O experimento foi conduzido para avaliar a eficiência de enzimas exógenas em dietas sobre o desempenho de frangos de corte. Um total de 1440 pintos de corte machos (Cobb 500®) foram distribuídos em um delineamento inteiramente casualizado em esquema fatorial 2x2 (duas dietas com diferentes densidades nutricionais; com e sem a adição enzimática) com 8 repetições de 45 aves em cada unidade experimental. As dietas com diferentes densidades nutricionais foram: controle positivo (níveis normais) e um controle negativo (com redução de 4,3 e 4,5% de energia metabolizável; 16,7 e 17,7% de cálcio e 35 e 42,7% de fósforo, nas fases inicial e de crescimento, respectivamente). A suplementação enzimática consistiu da combinação das enzimas fitase (100g t-1) e amilase, xilanase e protease (500g t-1). Foi avaliado o desempenho das aves nas fases de 1 a 21 e 1 a 42 dias de idade. Na fase total de criação, as aves alimentadas com a dieta controle negativo com adição enzimática tiveram consumo de ração (5,97%), peso médio (8,47%), ganho de peso (8,64%) e conversão alimentar (2,92%) melhores (P<0,05), comparados às aves alimentadas com dieta controle negativo, sem a suplementação de enzimas. Além disso, as aves alimentadas com a dieta controle negativo com adição enzimática apresentaram consumo de ração, peso vivo médio e ganho de peso similar (P>0,05) ao grupo alimentado com a dieta controle positivo com ou sem enzima. A adição de enzimas exógenas em dieta de frangos de corte com redução de energia metabolizável, cálcio e fósforo, proporciona um consumo de ração, peso vivo médio e ganho de peso similar a uma dieta com níveis nutricionais adequados.

Desempenho, composição da cama, das tíbias, do fígado e das excretas de frangos de corte alimentados com rações contendo fitase e baixos níveis de fósforo

Um experimento foi realizado para avaliar o desempenho e a composição da cama, das tíbias, do fígado e das excretas de frangos de corte em diversas fases utilizando-se diferentes níveis de fósforo disponível (Pd) e da enzima fitase. Foram utilizados 1.200 pintos machos de 1 dia de idade, da marca comercial Cobb, distribuídos em delineamento inteiramente casualizado, em esquema fatorial 4 × 3, composto de quatro níveis de fósforo disponível e três níveis de fitase: 0, 500 e 1.000 ftu/kg de ração, cada um com quatro repetições de 25 aves. Os níveis de fósforo disponível foram obtidos considerando quatro disponibilidades do fósforo total (Pt) do milho e do farelo de soja (34, 56, 78 e 100% de disponibilidade). Foram avaliados os índices de desempenho no final de cada fase de criação e aos 42 dias de idade. Os parâmetros de desempenho foram afetados pela redução dos níveis de fósforo disponível das rações e a redução desse elemento para níveis inferiores a 19% piorou o desempenho das aves, enquanto a utilização de fitase no nível de Pd reduzido em 39% melhorou o desempenho. Nas fases de criação, a redução do teor de fósforo disponível com a utilização da enzima fitase influencia o desempenho e as características da cama e dos ossos.

Estabilidade da fitase e sua utilização na alimentação de frangos de corte

This work was designed to evaluate the effect of storage forms and conditions upon the enzyme activity of phytase and bioavaibility of calcium and phosphorus in broiler diets. The work was accomplished in two steps. The first step, made in the laboratory measured the activity of the phytase enzyme along the storage period. In this step, two experiments were performed: Experiment 1, constituted of 5 treatments (pure phytase stored at 0 °C, 4 °C and environmental temperature and mixed to vitamin and mineral supplement, stored at environmental temperature) in CRD and split plot scheme. The activities were evaluated every 14 days for 112 days of storage, being verified that the phytase storage in the pure form at 0o C was superior to the other treatments. Experiment 2, made up of 4 treatments (phytase mixed to the ration directly, directly and afterwards pelleted, via mineral supplement and via vitamin supplement),all the treatments being stored at environmental temperature, in CRD and split plot scheme. The activities were evaluated every 7 days for 56 days' storage, being verified that the storage of the phytase mixed to the ration via vitamin supplement and directly with the ration pelleted later, provoked a fall in phytase activity when compared with the other treatments. In the second step, the effect of phytase on the bioavaibility of calcium and phytic phosphorus was evaluated, 2 experiments being accomplished (3 and 4); in both experiments were utilized 576 broiler line chicks, housed in an array of heated batteries, receiving practical diets on the basis of corn and soybean meal (basal) for 21days. At the end of 27 days of age,96 birds were slaughtered for evaluation of the mineral contents (Ca and P) in the tíbias and plasma phosphorus. The excretae were collected from 22 to 27 days of age of the birds. Experiment 3: A CRD with the treatments in 2 x 3 x 2 +4 factorial arrangement was utilized, namely, two levels of total phosphorus (0.35 and 0.45% of total phosphorus), three leveis of phytase (500, 750 and 1,000 FTU) and four additional treatments with levels of 0.35 and 0.45 % of available phosphorus for each sex, with three replicates per treatment. There was significant interaction among levels of phosphorus and phytase (P< 0.05) for weight gain, ration consumption and feed conversion. Phytase did not indicate significant differences when the level 0.45% was utilized, nevertheless, at the level 0.35% as phytase was supplemented, weight gain, ration consumption and feed conversion were improved, chiefly with 1,000 FTU/Kg, in both sexes. The males presented greater weight gain. The ration consumption and feed conversion were equal to those of females. The contrast 0.45% did not affect the performance of males and females, the same not occurring with the level 0.35%,at which the available phosphorus was superior in both the sexes. The highest contents of ashes, phosphorus and calcium in the tíbias and plasma phosphorus were obtained with the levels of 750 and 1,000 FTU/Kg of phytase and 0.45% of total phosphorus. The males presented higher contents of ashes in the tibias. The level 0.45% of available phosphorus presented the greatest contents of ashes, calcium and phosphorus in the tibias, and phosphorus in the plasma. The lowest excretions of phosphorus occurred at the levels 0.35% of total phosphorus and 1,000FTU/kg of phytase. The lowest contents of ashes and calcium in the excretae were obtained with 0.35% and 1,000FTU/Kg of phytase. The females excreted smallest amounts of ashes, calcium and phosphorus than the males. Experiment 4: a CRD with the treatments in 3 x 4 x 2 factorial arrangement, namely, three levels of phytase (0, 500 and 1,000 FTU), four levels of calcium (0.7, 0.8, 0.9 and 1.0%) with four replicates per treatment. The performance was not affected by the treatments utilized, the males being superior to the females in weight gain, feed consumption and conversion. The contents of ashes in the tibias were not affected by the levels of phytase but as calcium levels raised, the ash contents increased. The contents of calcium and phosphorus in the tibias increased with the supplementation of 500 and 1,000 FTU/kg of phytase and with calcium levels. The utilization of phytase did not decrease the excretion of ashes, calcium and phosphorus.

Probiótico e enzimas em dietas de frangos de corte: desempenho, características da cama e excretas e produção de biogás

Pós-graduação em Zootecnia - FCAV

Associação de carboidrases e fitase em dietas valorizadas e seus efeitos sobre desempenho e qualidade dos ovos de poedeiras leves

A total of 350 commercial Bovans White laying hens were used to evaluate the association of carbohydrases and phytase in enriched diets and its effects on performance and egg quality of laying hens. The experiment used a randomized design with five treatments and seven replicates. The treatments were: 1. Positive control without added enzymes and without nutrient enrichment, 2. Negative control (NC) 1 with 1.5% and 6% AME (kcal/kg) enrichment for corn and soybean meal respectively, 2% crude protein (CP) enrichment, and digestible limiting digestible amino acids plus the full matrix for the phytase enzyme; 3. NC 2 with 1.5% and 6% AME (kcal/kg) enrichment, respectively, for corn and soybean meal and 2% crude protein (CP) enrichment, and digestible limiting amino acids plus the sparse matrix for the phytase enzyme, 4. NC 1 supplemented with 100 g ton(-1) carbohydrase and 30g ton(-1) phytase, 5. NC 2 supplemented with 100 g ton(-1) carbohydrase and 30g ton(-1) phytase. According to the results, the positive control treatments, NC1 and NC2, with or without enzyme supplementation, showed guaranteed performance for feed intake, egg yield, weight, egg loss and shell quality.

BIOPROCESS OPTIMIZATION AND CHARACTERIZATION OF PHYTASE FROM AN ENVIRONMENTAL ISOLATE BACILLUS MCCB 242

A study was undertaken to isolate phytase producers from environment and to segregate the most highly efficient phytase producer and to develop a bioprocess technology for commercial application. During this process, a potential phytase producer Bacillus MCCB 242 was isolated and characterized phenotypically and genotypically. Subsequently, phytase production was optimized, the enzyme purified and characterized and an appropriate downstream process also could be standardized.Precisely, through this work an environmental isolate Bacillus MCCB 242 could be brought out as phytase producer for commercial application. The enzyme production could be optimized and characterized, and an appropriate downstream process standardized. Cytotoxicity studies revealed the enzyme safe for feed application, especially in fish.

Isolation and characterisation of phytase from dormant Corylus avellana seeds

Phytase (myo-inositol-1,2,3,4,5,6-hexakisphosphate phosphohydrolase, EC 3.1.3.26), which catalyses the step-wise hydrolysis of phytic acid, was purified from cotyledons of dormant Corylus avellana L. seeds. The enzyme was separated from the major soluble acid phosphatase by successive (NH4)2SO4 precipitation, gel filtration and cation exchange chromatography resulting in a 300-fold purification and yield of 7.5%. The native enzyme positively interacted with Concanavalin A suggesting that it is putatively glycosylated. After size exclusion chromatography and SDS–PAGE it was found to be a monomeric protein with molecular mass 72±2.5 kDa. The hazel enzyme exhibited optimum activity for phytic acid hydrolysis at pH 5 and, like other phytases, had broad substrate specificity. It exhibited the lowest Km (162 μM) and highest specificity constant (Vmax/Km) for phytic acid, indicating that this is the preferred in vivo substrate. It required no metal ion as a co-factor, while inorganic phosphate and fluoride competitively inhibited enzymic activity (Ki=407 μM and Ki=205 μM, respectively).

Carcass characteristics of broilers at 42 days receiving diets with phytase in different energy and crude protein levels

Objetivou-se, com este trabalho, estudar as características de carcaça e qualidade da carne do peito depois da inclusão de fitase em dietas para frangos de corte, com diferentes níveis de energia metabolizável aparente corrigida para nitrogênio (EMAn) e proteína bruta (PB) reduzida, suplementadas com aminoácidos essenciais seguindo o conceito de proteína ideal. Foram utilizados 1.500 frangos machos Cobb dos 22 aos 42 dias de idade com peso inicial de 833 ± 7 g e final de 2741 ± 48 g distribuídos em delineamento inteiramente casualizado em esquema fatorial 3x3+1 (três níveis de EMAn - 2950, 3100 e 3250 kcal/kg - e três de PB - 14, 16 e 18% - e um tratamento adicional - controle, sem fitase, com 3100 kcal/kg EMAn, 19,2% de PB e 0,4% de fósforo disponível) em seis repetições com 25 aves cada. Ao final do experimento, duas aves de cada parcela foram sacrificadas para a mensuração do rendimento de carcaça e de cortes e determinação da composição química da carne do peito. Os níveis de energia e proteína em rações com fitase influenciaram (P<0,05) os rendimentos de carcaça, peito e gordura abdominal a porcentagem de umidade, proteína e lipídios no músculo pectoralis major das aves, sendo os níveis de 3100 kcal EMAn/kg e 18% de PB os que proporcionaram maiores rendimentos de carcaça e de peito e menor deposição de gordura abdominal, mas em maior teor de lipídios na carne do peito. Conclui-se que a manipulação da energia em rações com reduzido teor de proteína e suplementadas com aminoácidos e fitase influencia o rendimento de cortes e a qualidade da carne do peito de frangos aos 42 dias.

Fabrication of phytic acid sensor based on mixed phytase-lipid Langmuir-Blodgett films

This paper reports the surface activity of phytase at the air-water interface, its interaction with lipid monolayers, and the construction of a new phytic acid biosensor on the basis of the Langmuir-Blodgett (LB) technique. Phytase was inserted in the subphase solution of dipalmitoylphosphatidylglycerol (DPPG) Langmuir monolayers, and its incorporation to the air-water interface was monitored with surface pressure measurements. Phytase was able to incorporate into DPPG monolayers even at high surface pressures, ca. 30 mN/m, under controlled ionic strength, pH, and temperature. Mixed Langmuir monolayers of phytase and DPPG were characterized by surface pressure-area and surface potential-area isotherms, and the presence of the enzyme provided an expansion in the monolayers ( when compared to the pure lipid at the interface). The enzyme incorporation also led to significant changes in the equilibrium surface compressibility (in-plane elasticity), especially in liquid-expanded and liquid-condensed regions. The dynamic surface elasticity for phytase-containing interfaces was investigated using harmonic oscillation and axisymmetric drop shape analysis. The insertion of the enzyme at DPPG monolayers caused an increase in the dynamic surface elasticity at 30 mN m(-1), indicating a strong interaction between the enzyme and lipid molecules at a high-surface packing. Langmuir-Blodgett (LB) films containing 35 layers of mixed phytase-DPPG were characterized by ultraviolet-visible and fluorescence spectroscopy and crystal quartz microbalance nanogravimetry. The ability in detecting phytic acid was studied with voltammetric measurements.