Dietary phytate induces subclinical mechanical allodynia in mice

Neuropathic pain is a condition with varying origins, including reduced dietary micronutrient intake. Phytate is a polyphosphate found in seeds and grains that can act as an antinutrient due to the ability of sequester essential divalent metals. Here we tested whether moderate dietary phytate intake could alter nociceptive pain. We subjected weaning mice to a chow supplemented with 1% phytate for eight weeks. Body weight gain, glycemic responses, food ingestion, water ingestion, and liver and adipose tissue weights were not altered compared to controls. We observed a decreased mechanical allodynia threshold in the intervention group, although there were no changes in heat- or cold-induced pain. Animals consuming phytate showed reduced spinal cord tumor necrosis factor (TNF), indicating altered inflammatory process. These data provide evidence for a subclinical induction of mechanical allodynia that is independent of phytate consumption in animals with otherwise normal phenotypic pattern.

Isolation of phosphate-solubilizing bacteria from subtropical soils with different fertilization histories / Isolamento de bactérias solubilizadoras de fosfato de solos subtropicais com diferentes histórias de fertilização

Phosphorus is one of the most abundant chemical elements but has a low bioavailability index. Therefore, microrganisms play a fundamental role in providing insoluble phosphorus to plants. The objective of this study was to evaluate the capacity of bacteria to solubilize inorganic phosphates in soils with different fertilization histories. Soil and rhizosphere samples were collected from a Red Distroferric Latosol, including a control without mineral or organic fertilizer (C), treatment with mineral fertilizer (MF) according to the needs of each crop, and treatment with organic fertilizer [300 m3 ha-1 of swine wastewater (SW)]. The medium containing calcium phytate presented more colony-forming units (CFU) for all fertilization histories, and growth in treatments C and MF was 50% higher than treatment with SW. CFU values in soils treated with SW were lower than those in the other treatments, and the diversity of insoluble phosphate-solubilizing bacteria (PSB) was higher in treatment C. These results indicate a negative relationship between phosphorus concentrations and the number of PSB.

O fósforo é um dos elementos químicos mais abundantes, mas tem um baixo índice de biodisponibilidade. Portanto, micro-organismos desempenham um papel fundamental no fornecimento de fósforo insolúvel para as plantas. O objetivo deste trabalho é avaliar a capacidade das bactérias em solubilizar fosfatos inorgânicos em solos com diferentes históricos de fertilização. Amostras de solo e rizosfera foram coletadas de um Latossolo Vermelho Distroférrico, incluindo controle sem adubação mineral ou orgânica (C), tratamento com adubação mineral (MF) de acordo com as necessidades de cada cultura e tratamento com adubação orgânica [300 m3 ha-1 de águas residuárias da suinocultura (SW)]. O meio contendo fitato de cálcio apresentou mais unidades formadoras de colônias (UFC) para todas as histórias de fertilização, e o crescimento nos tratamentos C e MF foi 50% maior que o tratamento com SW. Os valores de CFU nos solos tratados com SW foram menores que nos demais tratamentos, e a diversidade de bactérias insolúveis solubilizadoras de fosfato (PSB) foi maior no tratamento C. Esses resultados indicam uma relação negativa entre as concentrações de fósforo e o número de PSB.

Characterization and comparison of phytase production by Bacillus and Paenibacillus strains from Thai soils

Aims: The objective of this research was to isolate, screen and identify phytase-producing bacteria from soils and a potent isolate was selected for its phytase production. Methodology and results: Eight spore-forming bacteria isolated from agricultural soils in Thailand were screened for their phytase production. They were identified as Bacillus and Paenibacillus strains based on their phenotypic characteristics and 16S rRNA gene sequence analyses. The phytase production by Bacillus amyloliquefaciens CH3-1 [Group I(a)] was 20.956 ± 0.099 U/mL, while Bacillus subtilis SR9-3 [Group I(b)] produced 20.588 ± 0.099 U/mL. Five isolates in Group I(c), identified as Bacillus aryabhattai, produced phytase at levels ranging from 2.436 ± 0.116 to 20.910 ± 0.000 U/mL, while Paenibacillus cineris CM5-3 (Group II) produced 1.261 ± 0.111 U/mL. A potent strain, CH3-1, produced the highest phytase when cultivated in Phytate Specific Medium (PSM) supplemented with 1% glucose, at pH 7.0 and incubated at 45 °C. Additionally, wheat bran and sorghum seed (0.5%) substrates were used to induce phytase production by replacing Na-phytate. Conclusion, significance and impact of study: Phytase producing bacteria were isolated from soils in Thailand. Gram-positive spore forming thermotolerant Bacillus strains displayed higher phytase activity than a Paenibacillus strain. A potent strain, CH3-1, could utilize agricultural waste as a substrate, which may be useful for animal feed supplementation.

Adherence of Streptococcus pneumoniae and expression analysis of neuraminidase gene (NanA and NanB) after interaction of A549 human lung epithelial cells with pneumococcal strains of various serotypes

Aims: Streptococcus pneumoniae is the most common bacterial respiratory pathogen that can lead to invasive diseases such as pneumonia, bacteremia, and meningitis. The interaction of S. pneumoniae with host respiratory epithelial cells is crucial in the colonization of human respiratory tract and involve in the virulence. The aim of the study is to investigate the adherence of S. pneumoniae and the effect of serotypic variation on neuraminidase genes (NanA and NanB) after interaction of A549 human lung epithelial cells with S. pneumoniae serotypes. Methodology and results: Six different serotypes of S. pneumoniae were used (1, 3, 5, 19F, 23F, and 14). A549 human lung epithelial cells were inoculated with pneumococcal strains of different serotype for 3 hours. The number of adherent bacteria was determined by serial dilution followed by spread plate technique on tryptic soy agar supplemented with 5% sheep blood. Bacterial RNA was harvested from the infected A549 cells. The differential expression level of neuraminidases was observed by quantitative real-time PCR (qRT-PCR). Based on bacterial adherence assay, serotype 14 showed highest adherence, meanwhile, serotype 23F showed lowest adherence. This suggests that serotype 14 has a better affinity to adhere to A549 cells as compared to serotype 23F. Higher NanA gene expression was observed in serotype 5, 23F and 19F, while lower expression in serotype 14. In contrast, NanB gene shows low-level expression in serotype 23F and 19F, while higher expression in serotype 14. This postulates that NanA and NanB gene may have different functions in the pathogenesis of S. pneumoniae. Conclusion, significance and impact of study: Our finding on differential expression of neuraminidase gene of S. pneumoniae of various serotypes on A549 cells might give a better understanding of host pathogen interaction between bacteria serotypes and host cell.

Feasibility of sentinel lymph node dissection using Tc-99m phytate in papillary thyroid carcinoma

PURPOSE: Various methods of sentinel lymph node (SLN) biopsy in thyroid cancer have been introduced. Tc-99m phytate as a radiotracer has been successfully utilized for SLN biopsy in breast, cervix, and endometrial cancer. We assessed the feasibility of SLN dissection using Tc-99m phytate in papillary thyroid carcinoma (PTC). METHODS: Seventeen patients with PTC were prospectively enrolled. Ultrasound-guided peritumoral injection of 55.5 MBq Tc-99m phytate in 0.25-mL normal saline was performed. Preoperative single-photon emission-computed tomography (SPECT) and intraoperative gamma-probe were used for SLN detection during operation. RESULTS: Identification rate of SLNs was 70.6% (12 of 17) with SPECT, and 88.2% (15 of 17) with gamma-probe. Combined SPECT and gamma-probe had identification rates of 88.2% (15 of 17). Identification rates of SLNs in central LN compartments were 82.4% (14 of 17) and 41.2% (7 of 17) in lateral LN compartments. Overall sensitivity, specificity, positive predictive value, and negative predictive value of the results of SLN biopsy were 91.6%, 100%, 88.4%, and 100%, respectively. Eight patients (47.1%) had metastasis in SLNs; all patients had SLN metastasis in the central compartment and 2 patients had SLN metastasis in both the central and lateral compartments. CONCLUSION: Combined SPECT and gamma-probe could detect SLNs with an 88.2% identification rate in PTC. SLN biopsy using Tc-99m phytate is technically feasible. Further investigation is warranted for clinical application of Tc-99m phytate in PTC.

Contenido de calcio, fibra dietaria y fitatos en diversas harinas de cereales, pseudocereales y otros / Calcium, dietary fiber and phytate content in various flours of cereals, pseudocereals and other / Conteúdo de cálcio, fibra alimentar e fitatos em diversas farinhas de cereais, pseudocereais e outros

El contenido de calcio (Ca) y de compuestos que afectan su absorción son relevantes en dietas ricas en harinas integrales y frutos secos, tanto en vegetarianos estrictos, individuos con intolerancia a lácteos, como en celíacos. Se determinaron Ca, fibra dietaria y perfil de inositoles fosfato (IP) en harinas integrales/refinadas y/o extrudidas de cereales (trigo, maíz, arroz, sorgo), pseudocereales (trigo sarraceno, amaranto), leguminosas (arveja), oleaginosas (soja, lino). Se cuantificó Ca por espectrometría de absorción atómica, fibra dietaria por método enzimático-gravimétrico y perfil de IP por HPLC. Mediante factores de conversión, se calculó el contenido de ácido fítico (AF) y la relación molar AF/Ca como indicador de su disponibilidad potencial. El Ca (mg/100 g) fue muy bajo en harinas de cereales (<20 trigo refinada, maíz integral, arroz integral, sorgo integral y refinada), en pseudocereales fue variable (4 trigo sarraceno, 115 amaranto) y fue superior en harinas de arveja, soja y lino (102-257) pero no aumentó en galletitas con agregado de estas últimas harinas. El contenido de fibra dietaria fue acorde al grado de extracción (1,4-13,3 g/100 g). En todos los casos la relación molar AF/Ca superó 0,17-0,24, valor límite que disminuiría la absorción del Ca.

Calcium (Ca) content and compounds that affect its absorption are relevant in wholemeal flours and tree nuts. These foods are specially recommended for vegetarians, people with intolerance to dairy products and celiac patients. Ca, dietary fibre and inositol profile (IP) where analyzed in wholemeal/refined/ extruded flours, in cereals (wheat, maize, rice, sorghum), pseudocereals (buckwheat, amaranth), leguminous (peas) and oilseeds (soy, flax). Ca was assessed by atomic absorption spectrometry, dietary fibre using an enzymatic method and IP profile by HPLC. Phytic acid content (AF) was obtained using conversion factors. Molar ratio AF/Ca was calculated as an index of potential Ca availability. Ca (mg/100 g) was very low in cereal flours (<20 in refined wheat, wholemeal maize and rice and refined and wholemeal sorghum). In pseudocereals, Ca varied (4 buckwheat, 115 amaranth). It was high in peas, soy and flax (102-257), but this was not enough to improve Ca in cookies made with these mix flours. Dietary fibre content was coincidental with the extraction grade (1.4-13.3/100 g). In all cases, AF/Ca molar ratios exceeded 0.17-0.24, limit value that represents the negative effect of AF over calcium availability.

O teor de cálcio (Ca) e de compostos que afetam sua absorção são relevantes em dietas ricas em farinhas integrais e frutos secos, tanto em vegetarianos rigorosos, indivíduos com intolerância a laticínios e quanto celíacos. Determinaram-se Ca, fibra alimentar e perfil inositol fosfato (IP) em farinhas integrais/refinadas e/ ou extruidas de cereais (trigo, milho, arroz, sorgo), pseudocereais (trigo sarraceno, amaranto), leguminosas (ervilha), oleaginosas (soja, linhaça). Quantificou-se Ca por espectrometría de absorção atômica, fibra alimentar pelo método enzimático-gravimétrico e perfil de IP por HPLC. A través de fatores de conversão foi calculado o conteúdo de ácido fítico (AF) e a relação molar AF/Ca como indicador de sua disponibilidade potencial. O Ca (mg/100 g) foi muito baixo em farinhas de cereais (<20 trigo refinada, milho integral, arroz integral, sorgo integral e refinada), em pseudocereais foi variável (4 trigo sarraceno, 115 amaranto) e foi superior em farinhas de ervilha, soja e linhaça (102-257), mas não aumentou em biscoitos com estas últimas farinhas adicionadas. O conteúdo de fibra alimentar foi conforme ao grau de extração (1,4-13,3 g/100 g). Em todos os casos a relação molar AF/Ca superou 0,17-0,24, valor limite que diminuiria a absorção do Ca.

Phytase-Producing Bacteria from Extreme Regions in Indonesia

ABSTRACTIn this study, 154 isolates capable of producing extracellular phytate-degrading activity were isolated from four soil samples from volcanic areas in Central Java, Indonesia. Six strains with high phytate-degrading activity were selected for strain identification and characterization of the corresponding phytate-degrading enzyme. Blast analysis of 16S rRNA gene sequences revealed high similarities for all the six isolates to reference sequences belonging to the genusBacillus. Isolates MS5, MC6, D10 and D16 showed 99% sequence identity toB. cereus, while isolate MC8 exhibited 99% sequence identity toB. aryabhatti and D6 99% sequence identity toB. psychrotolerans. The crude extracellular phytase preparations from the isolates showed following optimal conditions for phytate dephosphorylation: pH 4.0 and 50°C (isolate D10), pH 5.0 and 60°C (isolate MC6, and isolate MS5), pH 6.0 and 50°C (isolate D16) and pH 6.0 and 60°C (isolate D6) and pH 6.0 and 40°C (isolate MC8). Zn2+ and Fe3+ strongly inhibited phytate dephosphorylation with all phytase preparations studied. In the presence of Ca2+, an increase in phytase activity of 10-15% was obtained.

Disponibilidade do fósforo em alimentos de origem vegetal: atuação na nutrição de peixes / Phosphorus availability in food from vegetable origin: performance in fish nutrition / Disponibilidad del fósforo en alimentos de origen vegetal: actuación en la nutrición de peces

O trabalho aborda a utilização do fósforo na nutrição de peixes e em dietas formuladas com ingredientes de origem vegetal. Nestes ingredientes, o fósforo encontra-se de forma indisponível para animais monogástricos, devido à carência da enzima fitase. A suplementação de enzimas exógenas (fitase) em ração tem por objetivo aumentar a capacidade de absorção do fósforo em dietas contendo ingredientes de origem vegetal, reduzindo a perda por excreção deste nutriente para o ambiente aquático. A enzima atua hidrolisando o fitato, tornando-o disponível para o organismo. Devido à importância do fósforo na nutrição de peixes, esta revisão apresenta alguns aspectos, sobre a importância na nutrição bem como sobre os fatores antinutricional e os benefícios causados pela inclusão da enzima em dietas balanceadas para peixes.

This paper discusses the use of phosphorus in fish nutrition and diets formulated with ingredients from vegetable origin. These ingredients from, phosphorus is unavailable for monogastric animals, due to the lack of the phytase enzyme. The supplementation of exogenous enzymes (phytase) in feed is intended to increase the phosphorus absorption capacity in diets containing ingredients from vegetable origin, reducing the loss of such nutrient by excretion into the aquatic environment. The enzyme acts hydrolyzing phytate, making it available to the body. Due to the importance of phosphorus in fish nutrition, this review presents some aspects importance, and anti-nutritional factor and benefits caused by the inclusion of the enzyme in fish balanced diets.

En este trabajo se aborda el uso de fósforo en la nutrición de peces y en dietas formuladas con ingredientes de origen vegetal. Estos ingredientes, el fósforo se encuentra de forma indisponible para animales monogástricos, debido a la falta de la enzima fitasa. La suplementación de enzimas exógenas (fitasa) en la ración tiene por objetivo aumentar la capacidad de absorción del fósforo en dietas que contienen ingredientes de origen vegetal, reduciendo la pérdida por excreción de este nutriente para el ambiente acuático. La enzima actúa hidrolizando el fitato, haciéndolo disponible para el cuerpo. Debido la importancia del fósforo en la alimentación de peces, esta revisión presenta algunos aspectos, tales como la importancia en la nutrición de peces, como el factor anti nutricional y los beneficios causados por la inclusión de la enzima en dietas balanceadas.

Dietas com fitase para frangos de corte alojados em ambientes com diferentes sistemas de climatização / Diets with phytase for broilers housed in environments with differenta air-conditioning systems

O experimento foi conduzido para avaliar o efeito de diferentes níveis de fitase nas dietas de frangos de corte sobre as variáveis de desempenho, rendimento de carcaça e cortes, incorporação de proteína bruta, cálcio e fósforo na carcaça e teores de nitrogênio, cálcio e fósforo na cama das aves aos 41 dias de idade. Seiscentos e quarenta pintos de um dia foram distribuídos em boxes em delineamento inteiramente casualizado, esquema fatorial 5 x 2, com cinco níveis de fitase: 0, 1000, 2000, 3000 e 4000 FTU por kg de ração, com quatro repetições e 16 aves na unidade experimental, associados a dois ambientes (galpão com ventilação e nebulização e galpão com ventilação sem nebulização). As aves receberam dietas isonutritivas à base de milho e farelo de soja, formuladas para atender as exigências nutricionais, exceto para fósforo disponível, que foi reduzido em 20% da exigência. O ambiente com ventilação e nebulização reduziu a temperatura média da pele, melhorou a viabilidade da criação e as variáveis de desempenho de frangos de corte. Além disso, aumentou a incorporação de proteína bruta e fósforo na carcaça das aves, bem como elevou os teores de nitrogênio, cálcio e fósforo na cama, em relação ao ambiente com apenas ventilação. A suplementação com fitase até 4000 FTU por kg da dieta aumentou o rendimento de peito.

The experiment was conducted to evaluate effect of different levels of phytase in broiler diets on performance variables, carcass yield and cuts, incorporation of crude protein, calcium and phosphorus in housing and levels of nitrogen, calcium and phosphorus in bed birds at 41 days of age. Six hundred and forty day-old chicks were distributed in boxes in a randomized design, factorial scheme 5 x 2, with five levels of phytase: 0, 1000, 2000, 3000 and 4000 FTU per kg diet, with four replications and 16 birds in experimental unit associated with two rooms (shed with ventilation and fogging and shed with ventilation without fogging). Birds were fed diets isonutrient maize and soybean meal, formulated to meet nutritional requirements, except for available phosphorus, which was reduced in 20% of requirement. The room with ventilation and fogging reduced skin temperature, improved feasibility and performance variables of broilers. It also increased incorporation of crude protein and phosphorus in poultry housing and elevated levels of nitrogen, calcium and phosphorus in bed, in relation to environment with only ventilation. Phytase supplementation to 4000 FTU per kg diet increased breast meat yield.

Cinética do P nos tecidos de suínos em crescimento alimentados com dietas formuladas de acordo com o conceito de proteína ideal e suplementadas com enzima fitase / Phosphorus kinetic in growing pigs tissue fed diets formulated according to the ideal protein concept and supplemented with phytase enzyme

O presente estudo foi desenvolvido para avaliar os efeitos da enzima fitase sobre a cinética do P nos tecidos de suínos, alimentados com dietas formuladas de acordo com o conceito de proteína ideal e suplementadas com enzima fitase. Foram utilizados 20 suínos machos castrados, distribuídos em um delineamento em blocos ao acaso, com cinco tratamentos e quatro repetições. Os animais foram alojados em gaiolas metabólicas durante 17 dias, sendo 10 dias para adaptação e sete dias para coletas de fezes e urina. Amostras de sangue foram coletadas por cinco dias. No primeiro dia da fase experimental, cada animal recebeu, por via endovenosa, uma solução radioativa com 7,4MBq de 32P. No final do período experimental, os animais foram sacrificados, e amostras de tecidos do músculo (lombo), coração, fígado, rins e ossos foram coletadas. A enzima fitase interferiu na cinética do 32P, levando a uma menor incorporação nos tecidos dos ossos...

This study was developed to evaluate the kinetic of the P in the pigs' tissues, feed diets formulated according the ideal protein concept and supplemented with phytase enzyme. Twenty male pigs were used, distributed in a randomized blocks experimental design, with five treatments and four replicates. The animals were housed in metabolic cages for 17 days, with 10 days for adaptation and 7 days for collection of feces and urine. Blood samples were collected in 5 days. On first day of the experimental phase, each animal was intravenously injected with 7.4 MBq 32P. At the end of the experiment the animals were slaughtered and samples of muscle (loin), heart, liver, kidney and bones were collected. The enzyme fitase interfered in the kinetics of 32P, leading to a smaller incorporation of bone tissues...

Increasing levels of phytase in diets formulated with reduced available phosphorus content supplied to male and female broilers / Níveis crescentes de fitase em dietas formuladas com reduzido teor de fósforo disponível, fornecidas a frangos de corte machos e fêmeas

The reduction of the available phosphorus (avP) content in the broiler diet must be followed by the supplementation of adequate phytase. One experiment was conducted with broilers from 15 to 28 days of age to determine the level of necessary phytase when the mashed diet is formulated with 0.213% of avP to enable the results of performance, tibia ash content, and calcium (Ca) utilization similar to those determined for birds fed with diet formulated to meet their nutritional requirements. Thus, 120 broilers were distributed in (4+1) x2 factorial arrangement corresponding to four deficient diets in avP (0.213%) supplemented with phytase (0; 750; 1,500 or 2,250FTU kg-1) plus one positive control diet without phytase (0.426% of avP), supplied to male and female broilers. Males showed better performance and higher totalP retention. Decrease in the avP content of the diet without phytase use worsened the performance, tibia ash content, and Ca retention; however, these parameters were improved gradually with the increase of the phytase level in the diet. Regardless of sex, using 2,250FTU kg-1, it is possible to reduce the avP to 0.213% without impairing performance, tibia ash content, and Ca retention; in addition to reducing the totalP excretion in 56.75% and improving its utilization in 38.58%.

A redução do teor de fósforo disponível (Pdisp) na dieta de frangos deve ser acompanhada de adequada suplementação da fitase. Um experimento foi conduzido com frangos de corte, no período de 15 a 28 dias de idade, para determinar o nível de fitase necessário quando a ração farelada é formulada com 0,213% de Pdisp para permitir resultados de desempenho, teor de cinzas na tíbia e aproveitamento do cálcio (Ca) semelhantes ao determinado para aves alimentadas com uma dieta formulada para atender suas exigências nutricionais. Desse modo, 120 frangos foram distribuídos em esquema fatorial (4+1) x2, correspondendo a quatro rações deficientes em Pdisp (0,213%) suplementadas com fitase (0; 750; 1.500 ou 2.250FTU kg-1) mais uma ração controle positivo sem fitase (0,426% de Pdisp), fornecidas a frangos machos e fêmeas. Os machos apresentaram melhor desempenho e maior retenção de Ptotal. A redução do teor de Pdisp da dieta sem o uso de fitase piorou o desempenho, o teor de cinzas ósseas e a retenção do Ca, todavia, esses parâmetros foram gradativamente melhorados com o aumento do nível de fitase na ração. Independente do sexo, ao utilizar 2.250FTU kg-1, é possível reduzir o teor de Pdisp para 0,213% sem prejudicar o desempenho, teor de cinzas na tíbia e a retenção do Ca, além de reduzir a excreção do Ptotal em 56,75% e melhorar seu aproveitamento em 38,58%.

Radioisótopo e modelagem matemática na avaliação da suplementação de fitase em dietas para suínos em crescimento / Radioisotope and mathematical modeling in the assessment of supplementation of Phytase in diets for growing pigs

O objetivo deste trabalho foi avaliar os efeitos da inclusão de níveis crescentes de enzima fitase em dietas para suínos em fase de crescimento, por meio do uso de modelagem matemática. Foram utilizados dados de 20 leitões mestiços, machos castrados, pesando em média, 26,8kg. Os animais ficaram alojados em gaiolas metabólicas individuais para a coleta de fezes e urina, onde permaneceram por um período de 17 dias. Utilizou-se o delineamento experimental de blocos ao acaso, com cinco tratamentos e quatro repetições. A dieta experimental fornecida aos leitões foi à base de milho e farelo de soja, suplementada com cinco níveis crescentes de enzima fitase (0, 250, 500, 750 e 1000UF/kg), correspondendo a 0; 0,01; 0,02; 0,03 e 0,04%, respectivamente. As variáveis avaliadas foram: ingestão, excreção, fluxo e refluxo do P nos compartimentos (trato digestivo, corrente sanguínea, tecidos moles e ossos). A enzima fitase não interferiu no P consumido (P>0,05 (F10), no P excretado na urina (F02), no fluxo e refluxo do P nos ossos (F32 e F23) e nos tecidos moles (F42 e F24), entretanto observou-se redução no P excretado nas fezes (F01) em 8,92%; 26,76%; 22,53% e 28,64% para os níveis 0, 250, 500, 750 e 1000UF/kg, respectivamente, e efeito linear positivo para o P endógeno (F12). Pode-se utilizar dietas à base de milho e farelo de soja com 50% de P por fosfato bicálcico, adicionando-se 250UF/kg de dieta para suínos em crescimento, o que reduz em 27% as excreções de P nas fezes.

The purpose of this study was to evaluate the effects of including increasing levels of phytase enzyme in pig diets for growing pigs, using the mathematical model. Data from 20 crossbred male piglets, castrated and weighing 26.80 kg on average was used. The animals were housed in individual metabolic cages to collect feces and urine in a 17 day period. A randomized block experimental design containing five treatments and four repetitions was used. The experimental diet provided to piglets contained corn and soybean and was supplemented with five increasing levels of phytase enzyme (0, 250, 500, 750 and 1000 UF/kg), corresponding to 0.00 %, 0.01 %, 0.02 %, 0.03 % and 0.04 % respectively. The variables evaluated were: intake, excretion, output flow of P in the digestive tract, bloodstream, bones and soft tissues. The phytase enzyme did not affect the P intake (P>0.05 (F 10), the P excreted in urine (F02) and the output flow of P in the bones (F32 e F23) and soft tissue (F42 e F24) , however, there was a reduction in P excreted in feces (F01) of 8.92 %, 26.76 %, 22.53 % and 28.64 % to the levels 0, 250, 500, 750 e 1000UF/kg, respectively and showed a positive linear effect (P<0.08) for the endogenous P (F12) . Corn and soybean meal based diets can be used with 50% of P by dicalcium phosphate, adding 250UF/kg diet for growing pigs, and may cause a reduction of 27 % of P excretion in feces.

Disponibilidad de fósforo soluble en ensilado experimental fermentado con una cepa de Lactococcus lactis subsp. lactis con alta actividad de fitasa / Soluble phosphorous availability in experimental silage fermented with a Lactococcus lactis subsp. lactis strain with high phytase activity

El fitato es la forma principal de almacenamiento de fósforo en semillas y granos, pero el fósforo unido a fitato resulta inaccesible para los peces. El objetivo del presente trabajo fue estudiar la eficacia de Lactococcus lactis subsp. lactis Tw34 para liberar fósforo de fitato en un alimento experimental que contenía harina de cereal. La bacteria inoculante del ensilado biológico se seleccionó por su alta actividad de fitasa y las mezclas se prepararon con productos de descarte de merluza (Merluccius hubbsi). Las harinas de girasol, maíz, trigo y cebada se mezclaron con desechos picados de merluza hasta alcanzar el 25% y se inocularon con la bacteria citada. Después de 7 días de fermentación a 18 ºC los mejores resultados se obtuvieron cuando se utilizó harina de cebada. La cepa fue capaz de reducir el valor del pH a 4,4 y aumentar cuatro veces la concentración inicial de fósforo libre. Los resultados sugieren que la fitasa de Lactococcus lactis subsp. lactis Tw34 puede aumentar la accesibilidad del fósforo en dietas basadas en harina de cereales, evitando los efectos negativos del ácido fítico en la biodisponibilidad de minerales y proteínas, y disminuyendo la liberación de residuos de fósforo.

Phytate is the main phosphorous storage form in grains and seeds, but phytate linked phosphorous is inaccessible for fishes. The purpose of this wok was to study the efficacy of Lactococcus lactis subsp. lactis Tw34 for liberating phosphorous in an experimental feed which contained cereal flour. The inoculating bacterium of the biological silage was selected due to its high phytase activity, and the mixtures were prepared with waste hake products (Merluccius hubbsi). Sunflower, corn, wheat and barley flours were mixed with waste ground hake products up to 25% and inoculated with the bacterium mentioned. After 7 days fermentation at 18 oC, the best results were obtained when barley flour was used. The strain was able to reduce the pH value to 4.4 and increase four times the initial concentration of free phosphorous. The results suggest that Lactococcus lactis subsp. lactis Tw34 can increase phosphorous accessibility in cereal flour based diets, avoiding the negative effects of phytic acid on the bioavailability of minerals and proteins, and decreasing the liberation of phosphorous residues.

Fórmula de soja - de heroína a vilã? Análise crítica ao artigo / Soy Protein Infant Formulae and Follow-On Formulae: A Commentary by the ESPGHAN Committee on Nutrition

As autoras apresentam comentários sobre artigo publicado pelo Comitê de Nutrição da Sociedade Europeia de Gastroenterologia, Hepatologia e Nutrição, a respeito do papel da soja na nutrição infantil e das recomendações sobre as alternativas que se apresentam ao pediatra no que diz respeito à alimentação do lactente com alergia ao leite de vaca...

Inhibitory Effects of Phytate to Bioavailability of Calcium, Ferum and Zinc in Local Soy Products / Jurnal Sains Kesihatan Malaysia

This study was carried out to determine the inhibitory effects of phytate on the bioavalaibility of calcium (Ca), ferum (Fe) dan zink (Zn) in selected soy-based products. Phytate exerts its inhibitory effects on the absorption of minerals such as Ca, Fe and Zn by forming insoluble and undigestible complexes. The inhibitory effect of phytate on the bioavailability of minerals like Ca, Fe and Zn was estimated by measuring their molar ratios. Phytate content was determined by using anion-exchange method while mineral contents were determined by Atomic Absorption Spectrophotometer (AAS). Soybased products were bought from the night markets and food stall located at Cheras and Sri Rampai, Kuala Lumpur. Each samples from both location was analysed in duplicates. Results from this study showed that soy cake and all dishes containing tempe did not contain phytate content. This may due to the presence of R. oligosporus that produces phytase and hydrolysed the phytate content during the fermentation process of tempe. There were fi ve food samples that have a phytate/ Ca molar ratio > 0.24, ten food samples with a phytate/ Fe molar ratio > 1, eight food samples have a phytate/ Zn molar ratio >15 and one food sample has a [phytate x Ca]/ Zn molar ratio > 200. The result of molar ratio phytate/ mineral in this study estimated the low bioavailability of Ca, Fe and Zn. In conclusion, soy cake and all dishes containing tempe contained high bioavailability of minerals, while tau-hoo pok contained low bioavailability of minerals

Isolation, morphological and molecular characterization of phytate-hydrolysing fungi by 18S rDNA sequence analysis

Phytate is the primary storage form of phosphate in plants. Monogastric animals like poultry, pigs and fishes have very low or no phytase activities in their digestive tracts therefore, are incapable to efficiently utilize phytate phosphorus from the feed. Phytase from microbial sources are supplemented to feedstuff of these to increase the uptake of phytate phosphorus. In the present work efforts were made to isolate and characterize proficient phytase producing fungi from soil. Phytase producing fungi were isolated using phytate specific medium. Fungal isolates were selected according to their higher phytase activities. These isolates were further characterized and identified by morphological and microscopic analysis and confirmed by amplification of 18S rRNA gene, using specific primers. This gene was subsequently sequenced and phylogenetic affiliations were assigned. Fungal isolates were identified as various species of Aspergillus. Phytases from these fungi could be utilized as a feed additive in poultry and swine industries.

Purification, characterization and properties of phytase from Shigella sp. CD2.

Phytases catalyze the release of phosphate from phytic acid. In this study, a phytase producing bacterial strain Shigella sp. CD2 was isolated from the wheat rhizosphere. Phytase production started from the exponential phase of bacterial growth, showing the highest activity during the stationary phase. The enzyme activity was detected in both periplasmic and intracellular fractions. The enzyme was purified by about 133-fold with specific activity 780 U mg-1 protein. The optimum pH and temperature of the enzyme was 5.5 and 60oC, respectively. The enzyme was thermostable and retained 100% and 75% of its activity on pre-incubation at 70o and 80oC for 30 min, respectively. The Km value for the substrate sodium phytate was 0.25 mM. The enzyme was highly specific to substrate phytate, and no activity was detected in presence of other phosphorylated substrates, such as ATP, ADP, glucose 6-phosphate, fructose 6-phosphate and p-nirophenyl phosphate. The activity declined dramatically in presence of Cu2+, Zn2+ and Fe2+ and SDS, whereas Mg2+ and Co2+ slightly enhanced the enzyme activity. The addition of other metal ions or chemicals had little or no effect on phytase activity. The enzyme was resistant to both pepsin and trypsin. Due to high specific activity, substrate specificity, good pH profile, protease insensitivity and thermostability, phytase encoding gene from Shigella sp. CD2 could be an interesting candidate for industrial applications. Further studies on cloning and expression of Shigella phytase gene are currently in progress.

Efficiency of phosphorus utilization in phyA-expressing cotton lines.

To evaluate and characterize the stability of traits conferred by phyA from Aspergillus ficuum, we examined expression of phyA in sexually-derived transgenic cotton progeny and assessed the capacity for phytate-utilization in T4 progeny. The gene (phyA) was expressed only in the roots, but not in the stem and leaf tissues. Phytase activity was 2.38-fold higher in transgenic line L2 than in wild-type (WT) plants. The amount of phosphorus in the leaves was also significantly higher in transgenic lines L2, L6 and L9. Among those lines, L2 performed best, showing increase in phosphorus contents of 10.67% (seedling stage), 8.78% (squaring), 11.77% (flowering) and 11.59% (boll-opening). The level of available phosphorus in the rhizosphere was 19% higher in soil containing transgenic plants compared with the WT. When both transgenic and WT were grown in the same type of soil, the number of bacteria, fungi and actinomycetes was not obviously different. These results indicated that although the T4 generation enhanced the utilization of phytate phosphorus, it had no influence on the number of soil microorganisms. Our findings also demonstrated that phytase-expression lines could be used for developing new varieties of cotton having improved phosphorus uptake from the soil.

Activity of fungal phytases stored in two ways in response to the period of storage at room temperature / Atividade de fitases fúngicas armazenadas de duas maneiras em resposta ao período de armazenamento à temperatura ambiente

Two distinct experiments were conducted simultaneously with phytases of Aspergillus oryzae and A. niger for determining enzyme activity in response to storage period (up to 180 days at room temperature - RT). In the first experiment, enzymes were stored as they were acquired (pure form) and the activity was measured periodically during 180 days of storage at RT. In the second experiment, the phytases were incorporated to a supplement containing vitamins, minerals, and amino acids; and then this supplement was stored at RT up to 180 days, so that every 30 days of storage was collected one aliquot from each replicate for determining enzymatic activity. In conclusion, the phytase activity is affected by storage duration. To ensure 80 percent of the initial activity, the phytases of A. oryzae and A. niger can be stored in the pure forms for up to 53 and 135 days at RT, respectively. However, if the phytases of A. oryzae and A. niger are incorporated to a supplement containing vitamins, minerals, and amino acids then the storage period at RT should not exceed 67 and 77 days, respectively.

Dois experimentos distintos foram realizados simultaneamente com fitases de Aspergillus oryzae e A. niger para a determinação da atividade enzimática em resposta ao período de armazenamento (por até 180 dias em temperatura ambiente - TA). No primeiro experimento, as enzimas foram armazenadas como adquiridas (forma pura) e a atividade foi determinada periodicamente durante 180 dias de armazenamento em TA. No segundo experimento, as fitases foram incorporadas a um suplemento contendo vitaminas, minerais e aminoácidos. Então este suplemento foi armazenado em TA por até 180 dias, de modo que, a cada 30 dias de armazenamento, uma alíquota de cada repetição foi coletada para a determinação da atividade enzimática. Conclui-se que a atividade da fitase é afetada pela duração do armazenamento. Para assegurar 80 por cento da atividade inicial, as fitases de A. oryzae e A. niger podem ser armazenadas nas formas pura por até 53 e 135 dias em TA, respectivamente. Entretanto, se as fitases de A. oryzae e A. niger são incorporadas a um suplemento contendo vitaminas, minerais e aminoácidos, o período de armazenamento em TA não deve exceder 67 e 77 dias, respectivamente.

Nitrogen, calcium and phosphorus balance of broilers fed diets with phytase and crystalline amino acids / Balanço de nitrogênio, cálcio e fósforo em frangos de corte recebendo dietas com fitase e aminoácidos cristalinos

The objective of this study was to evaluate the effect of reduced-crude protein (CP) and available phosphorus (aP) diets, added with phytase and amino acids for broilers in the growing (14 to 21 days old) and retirement (35 to 42 days) phases on nitrogen, potassium and phosphorus balance. Ninety Cobb broilers were distributed in a completely randomized design with three treatments and six replications with five and three birds per experimental unit in the growing and retirement phases, respectively. Diets were elaborated to contain reduced CP and aP levels (19 and 17 percent CP in the growing phase, with 0.34 percent aP; and 18 and 16 percent CP in the retirement phase, with 0.30 percent aP), supplemented with crystalline amino acids and 500 FTU of phytase/kg diet. These were compared to a control diet (21 percent CP and 0.46 percent aP for growing phase and 19 percent CP and 0.40 percent aP for retirement phase). The reduction of CP in four percentual units reduced the N excretion and increased the retention of this element, showing that the phytase had no influence on N balance when used the crystalline amino acids in the diets. The phytase reduced the P excretion and increased the retention ones. The potassium excretion decreased in reduced-CP diets, but this excessive reduction decreased the retention of this element, except in the retirement phase. It was concluded that the reduced-CP and aP diets, supplemented with phytase and amino acids, are efficient in reducing the pollutant power of the broilers excrements in the growing and retirement phases of production.

Objetivou-se, com este trabalho, avaliar o efeito de rações com reduzidos teores de proteína bruta (PB) e fósforo disponível (Pd), suplementadas com fitase e aminoácidos, para frangos de corte na fase de crescimento (14 a 21 dias) e terminação (35 a 42 dias de idade) sobre o balanço de nitrogênio, potássio e fósforo. Foram utilizados 90 pintos Cobb, distribuídos em um delineamento inteiramente casualizado com três tratamentos e seis repetições, com cinco e três aves por unidade experimental nas fases inicial e de crescimento, respectivamente. As rações foram formuladas para conterem níveis reduzidos de PB e Pd (19 e 17 por cento de PB na fase de crescimento, com 0,34 por cento de Pd e 18 e 16 por cento de PB na fase de terminação, com 0,30 por cento de Pd) e suplementadas com aminoácidos e 500 FTU de fitase/kg de ração Essas rações foram, então, que foram comparadas a uma ração controle (21 por cento de PB e 0,46 por cento de Pd para fase de crescimento e 19 por cento de PB e 0,40 por cento de Pd para fase de terminação). A redução da PB em quatro unidades percentuais reduziu a excreção, e aumentou a retenção de nitrogênio, mostrando que a fitase não interferiu no balanço de nitrogênio ao serem incluídos aminoácidos cristalinos nas rações. A excreção de potássio diminuiu em dietas com PB reduzida, mas esta redução em excesso diminuiu a retenção deste elemento, exceto na fase de retirada. Conclui-se que rações contendo níveis reduzidos de PB, cálcio e fósforo, suplementadas com fitase e aminoácidos, são eficientes em diminuir o poder poluente das excretas de frangos de corte.