Efeito prolongado da toxina botulínica associada à suplementação com zinco e fitase / Prolonged effect of botulinum toxin associated with zinc and phytase supplementation / Efecto prolongado de la toxina de botulino asociada con el suplemento de zinc y la fitasa

A toxina botulínica é produzida naturalmente pela bactéria anaeróbia gram- positiva Clostridium botulinum, sendo considerada uma neurotoxina com efeitos capazes de reduzir atividades do sistema nervoso parassimpático, principalmente em relação as desordens de movimentos. A alta efetividade e o resultado do efeito de bloqueio na contração da musculatura facial justificam sua aplicação regular na estética, atuando no tratamento e prevenção das rugas e linhas de expressão. Entretanto, para a ação da toxina botulínica é necessário que o Zinco (Zn) exerça a ação no bloqueio da contração muscular através do mecanismo da clivagem no receptor SNAP-25. O objetivo deste trabalho é investigar a correlação do mecanismo de ação da toxina botulínica no bloqueio da contração muscular associado com a suplementação via oral de cápsulas de zinco e fitase, para comprovar a efetividade na durabilidade de bloqueio muscular com TBA. A pesquisa foi realizada através da aplicação de um questionário aleatório para participantes da pesquisa que fazem aplicação de TBA na musculatura facial de ambos os gêneros, com idade acima de 18 anos e que realizam a suplementação prévia de Zn e Fitase para aumentar a durabilidade do efeito da neurotoxina. Em relação aos resultados encontrados observou-se uma mudança significativa na duração do efeito da TBA quando associado a suplementação prévia de Zn e fitase. Dos 44 participantes da pesquisa 22,7% responderam que ficaram satisfeitos com os efeitos da TBA sem suplementação prévia, enquanto 93,2% afirmaram no questionário uma duração maior do bloqueio muscular quando suplementaram previamente com Zn e Fitase. Conclui-se que a suplementação de Zn e fitase previamente a aplicação da TBA tem resultado positivo na durabilidade do bloqueio da contração da musculatura facial, o que desperta grande interesse na área da estética aplicada, sugerindo novas terapias complementares que possam assegurar a satisfação dos pacientes que são adeptos a este procedimento estético.

Botulinum toxin is produced naturally by the anaerobic gram-positive bacterium Clostridium botulinum, being considered a neurotoxin with effects capable of reducing activities of the parasympathetic nervous system, mainly in relation to movement disorders. The high effectiveness and the result of the blocking effect in the contraction of the facial musculature justify its regular application in aesthetics, acting in the treatment and prevention of wrinkles and lines of expression. However, for the action of botulinum toxin it is necessary that Zinc (Zn) exerts the action in blocking muscle contraction through the cleavage mechanism in the SNAP-25 receptor. The objective of this work is to investigate the correlation of the mechanism of action of botulinum toxin in the blocking of muscle contraction associated with oral supplementation of zinc capsules and phytase, to prove the effectiveness in the durability of muscle block with TBA.The research was carried out by applying a random questionnaire to research participants who apply TBA in the facial musculature of both genders, aged over 18 years and who perform the previous Zn and Fitase supplementation to increase the durability of the effect of neurotoxin. In relation to the results found, a significant change in the duration of the effect of TBA was observed when associated with previous Zn and phytase supplementation. Of the 44 participants in the survey, 22.7% responded that they were satisfied with the effects of ABR without prior supplementation, while 93.2% stated in the questionnaire a longer duration of muscle block when they had previously supplemented with Zn and Fitase. It is concluded that the Zn and phytase supplementation prior to the application of the TBA has positive result in the durability of the blockage of the contraction of the facial musculature, which arouses great interest in the area of applied aesthetics, suggesting new complementary therapies that can ensure the satisfaction of patients who are adherents to this aesthetic procedure.

La toxina botulínica es producida naturalmente por la bacteria anaeróbica Gram-positiva Clostridium botulinum y se considera una neurotoxina con efectos capaces de reducir las actividades del sistema nervioso parasiinfático, principalmente en relación con los trastornos del movimiento. La alta eficacia y el resultado del efecto bloqueante sobre la contracción de los músculos faciales justifican su aplicación regular en estética, actuando en el tratamiento y prevención de arrugas y líneas de expresión. Mientras tanto, para la acción de la toxina botulínica, es necesario que Zinc (Zn) ejerza la acción de bloqueo de la contracción muscular por el mecanismo de escisión del receptor SNAP-25. El objetivo de este trabajo es investigar la correlación del mecanismo de acción de la toxina botulínica en el bloqueo de la contracción muscular asociada a la suplementación oral de cápsulas de zinc y fitasa, para probar la eficacia de la durabilidad del bloqueo muscular con TBA. La investigación se realizó aplicando un cuestionario aleatorio a los participantes en la investigación que aplican TBA a los músculos faciales de ambos géneros mayores de 18 años y que realizan el suplemento previo de Zn y Fitase para aumentar la durabilidad del efecto neurotoxina. En relación con los resultados hallados, se observó un cambio significativo en la duración del efecto del TBA cuando se asoció con el suplemento de Zn y fitasa. De los 44 participantes en la investigación, el 22,7% respondió que estaban satisfechos con los efectos de la TBA sin suplementos previos, mientras que el 93,2% declaró en el cuestionario una larga duración del bloqueo muscular cuando se les había suplementado previamente con Zn y Fitase. Se concluye que la suplementación con Zn y fitasa previa a la aplicación del TBA ha producido un resultado positivo en la durabilidad del bloqueo de contracción muscular facial, que despierta gran interés en el área de estética aplicada, sugiriendo nuevas terapias complementarias que puedan garantizar la satisfacción de los pacientes que son adeptivos a este procedimiento estético.

Efeito prolongado da toxina botulínica associada à suplementação com zinco e fitase / Prolonged effect of botulinum toxin associated with zinc and phytase supplementation / Efecto prolongado de la toxina de botulino asociada con el suplemento de zinc y la fitasa

A toxina botulínica é produzida naturalmente pela bactéria anaeróbia gram- positiva Clostridium botulinum, sendo considerada uma neurotoxina com efeitos capazes de reduzir atividades do sistema nervoso parassimpático, principalmente em relação as desordens de movimentos. A alta efetividade e o resultado do efeito de bloqueio na contração da musculatura facial justificam sua aplicação regular na estética, atuando no tratamento e prevenção das rugas e linhas de expressão. Entretanto, para a ação da toxina botulínica é necessário que o Zinco (Zn) exerça a ação no bloqueio da contração muscular através do mecanismo da clivagem no receptor SNAP-25. O objetivo deste trabalho é investigar a correlação do mecanismo de ação da toxina botulínica no bloqueio da contração muscular associado com a suplementação via oral de cápsulas de zinco e fitase, para comprovar a efetividade na durabilidade de bloqueio muscular com TBA. A pesquisa foi realizada através da aplicação de um questionário aleatório para participantes da pesquisa que fazem aplicação de TBA na musculatura facial de ambos os gêneros, com idade acima de 18 anos e que realizam a suplementação prévia de Zn e Fitase para aumentar a durabilidade do efeito da neurotoxina. Em relação aos resultados encontrados observou-se uma mudança significativa na duração do efeito da TBA quando associado a suplementação prévia de Zn e fitase. Dos 44 participantes da pesquisa 22,7% responderam que ficaram satisfeitos com os efeitos da TBA sem suplementação prévia, enquanto 93,2% afirmaram no questionário uma duração maior do bloqueio muscular quando suplementaram previamente com Zn e Fitase. Conclui-se que a suplementação de Zn e fitase previamente a aplicação da TBA tem resultado positivo na durabilidade do bloqueio da contração da musculatura facial, o que desperta grande interesse na área da estética aplicada, sugerindo novas terapias complementares que possam assegurar a satisfação dos pacientes que são adeptos a este procedimento estético.

Botulinum toxin is produced naturally by the gram-positive anaerobic bacterium Clostridium botulinum, and is considered a neurotoxin with effects capable of reducing activities of the parasympathetic nervous system, especially in relation to movement disorders. The high effectiveness and the result of the blocking effect on the contraction of facial muscles justify its regular application in aesthetics, acting in the treatment and prevention of wrinkles and expression lines. However, for the action of botulinum toxin it is necessary that the Zinc (Zn) exerts the action of blocking muscle contraction through the cleavage mechanism in the SNAP-25 receptor. The objective of this study is to investigate the correlation of the mechanism of action of botulinum toxin in blocking muscle contraction associated with oral supplementation of zinc capsules and phytase, to prove the effectiveness in the durability of muscle blockade with TBA. The research was carried out by applying a randomized questionnaire to research participants who apply TBA to the facial muscles of both genders, over 18 years of age, and who perform prior supplementation of Zn and Phytase to increase the durability of the neurotoxin effect. In relation to the results found it was observed a significant change in the duration of the effect of TBA when associated with prior supplementation of Zn and phytase. Of the 44 research participants 22.7% responded that they were satisfied with the effects of TBA without prior supplementation, while 93.2% stated in the questionnaire a longer duration of muscle blockade when previously supplemented with Zn and Phytase. We conclude that the supplementation of Zn and phytase previously to the application of TBA has a positive result in the durability of the blockade of facial muscle contraction, which arouses great interest in the area of applied esthetics, suggesting new complementary therapies that can ensure the satisfaction of patients who are adept at this esthetic procedure.

La toxina botulínica es producida naturalmente por la bacteria anaeróbica Gram-positiva Clostridium botulinum y se considera una neurotoxina con efectos capaces de reducir las actividades del sistema nervioso parasiinfático, principalmente en relación con los trastornos del movimiento. La alta eficacia y el resultado del efecto bloqueante sobre la contracción de los músculos faciales justifican su aplicación regular en estética, actuando en el tratamiento y prevención de arrugas y líneas de expresión. Mientras tanto, para la acción de la toxina botulínica, es necesario que Zinc (Zn) ejerza la acción de bloqueo de la contracción muscular por el mecanismo de escisión del receptor SNAP-25. El objetivo de este trabajo es investigar la correlación del mecanismo de acción de la toxina botulínica en el bloqueo de la contracción muscular asociada a la suplementación oral de cápsulas de zinc y fitasa, para probar la eficacia de la durabilidad del bloqueo muscular con TBA. La investigación se realizó aplicando un cuestionario aleatorio a los participantes en la investigación que aplican TBA a los músculos faciales de ambos géneros mayores de 18 años y que realizan el suplemento previo de Zn y Fitase para aumentar la durabilidad del efecto neurotoxina. En relación con los resultados hallados, se observó un cambio significativo en la duración del efecto del TBA cuando se asoció con el suplemento de Zn y fitasa. De los 44 participantes en la investigación, el 22,7% respondió que estaban satisfechos con los efectos de la TBA sin suplementos previos, mientras que el 93,2% declaró en el cuestionario una larga duración del bloqueo muscular cuando se les había suplementado previamente con Zn y Fitase. Se concluye que la suplementación con Zn y fitasa previa a la aplicación del TBA ha producido un resultado positivo en la durabilidad del bloqueo de contracción muscular facial, que despierta gran interés en el área de estética aplicada, sugiriendo nuevas terapias complementarias que puedan garantizar la satisfacción de los pacientes que son adeptivos a este procedimiento estético.

Sunflower meal with and without phytase supplementation in diets for silver catfish (Rhamdia quelen) juveniles / Harina de girasol con y sin suplementación de fitasa en dietas para juveniles de bagre (barbudo, Col.; Rhamdia quelen) / Farelo de girassol com e sem suplementação de fitase em dietas para juvenis de jundiá ( Rhamdia quelen)

Abstract Background: The cost of feed in fish farming can exceed 70% of the total costs. Thus, the feed industry have sought to formulate balanced diets including low-cost by-products. Objective: To evaluate sunflower meal as a partial substitute (0, 10, and 20%) for soybean meal, with or without phytase supplementation (1,500 FTU/kg) in isoproteic (27% crude protein (CP)) and isoenergetic diets (3,400 kcal/ED/kg of feed) for silver catfish (Rhamdia quelen) juveniles. Methods: A total of 360 fish (weight: 15.77 ± 0.56 g) were randomly distributed in 24 tanks in a completely randomized design following a factorial scheme (3x2) with 4 replicates. The fish were evaluated on the following variables after 90 days: productive performance, hematological variables, histology of intestinal tissue, bromatological composition of carcass, and composition of bone mineral matter. Results: 20% dietary inclusion of sunflower meal improved weight gain and protein efficiency. No difference was observed in the other treatments and no interactions were observed between the other factors. Conclusion: 20% sunflower meal can be included in the feed of silver catfish as a substitute for soybean meal. Phytase inclusion (1,500 FTU/kg) did not improve the nutritional value of the treatments.

Resumen Antecedentes: El costo de la alimentación en piscicultura pueden superar el 70% del costo total. Las industrias de alimentos para peces han intentado formular dietas balanceadas con subproductos de menor valor en el mercado para minimizar costos. Objetivo: Evaluar la harina de girasol como sucedáneo parcial (0, 10 y 20%) de la torta de soya, suplementada o no con fitasa (1.500 FTU/kg) en dietas isoprotéicas (27% de PB) e isoenergéticas 3400 kcal / ED / kg de ración) para bagre (barbudo, Col.; Rhamdia quelen). Métodos: Los peces, 360 juveniles (15,77 ± 0,56 g), se distribuyeron aleatoriamente en 24 unidades experimentales, en un diseño completamente al azar, en esquema factorial (3x2) y 4 repeticiones. A los 90 días se evaluó: desempeño productivo, variables hematológicas, histología de tejido intestinal y composición bromatológica de la carcasa y de la materia mineral de los huesos. Resultados: La inclusion de 20% de harina de girasol mejoró la ganancia de peso de los peces y la tasa de eficiencia proteica de la ración, sin diferencia para las otras evaluaciones ni interacción entre los demás factores evaluados. Conclusión: La harina de girasol se puede utilizar en un 20% de la ración del bagre (barbudo, Col.) como sucedáneo a la harina de soja. La inclusión de fitasa (1.500 FTU/kg) no mejoró el valor nutritivo de estas raciones.

Resumo Antecedentes: Os custos relativos a alimentação na piscicultura pode ultrapassar 70% do custo final, neste sentido, as indústrias de rações têm procurado formular dietas balanceadas com subprodutos de menor valor de mercado para minimizar estes custos. Objetivo: Esse estudo objetivou avaliar o farelo de girassol como sucedâneo parcial (0, 10 e 20%) do farelo de soja, suplementado ou não com a fitase (1.500 FTU/kg), em dietas isoprotéicas (27% de PB) e isoenergéticas (3400 kcal/ED/kg de ração) para o jundiá (Rhamdia quelen). Métodos: Os peixes, 360 juvenis (15,77 ± 0,56 g) foram distribuídos aleatoriamente em 24 unidades experimentais, num delineamento inteiramente casualizado, em esquema fatorial (3x2) e 4 repetições. Após 90 dias foram avaliados: desempenho produtivo, variáveis hematológicas, histologia de tecido intestinal, composição bromatológica da carcaça e de matéria mineral dos ossos. Resultados: O emprego de 20% de farelo de girassol melhorou o ganho de peso dos peixes e a taxa de eficiência protéica da ração, sem diferença nas outras avaliações, ou para a interação entre os demais fatores avaliados. Conclusão: Pode-se concluir que o farelo de girassol pode ser utilizado em 20% da ração do jundiá, como sucedâneo ao farelo de soja e, que a inclusão de fitase (1,500 FTU/kg) não melhorou o valor nutritivo destas rações.

Effect of Microbial Phytase on Nutrient Availability and Growth of Juvenile Clarias gariepinus Fed Soyabean and Groundnut-based Diets

Several studies have shown the positive effect of phytase on phosphorus utilization by fish and animals, with the use of phytase sources determined for different fish species. Few studies have tested phytase response to different diets, which may affect nutrient availability for optimum growth due to differences in phytate location. The research, therefore, studied the effect of phytase to diets based on soya bean and groundnut meal for Clarias gariepinus on nutrient availability and growth. In trial 1, four groups of soya bean S1, S2, S3 and S4 replaced fish meal at 25%, 50%, 75% and 100% and supplemented with 250, 500, 750 and 1000 FTU/g phytase, respectively. In trial 2, four groups of groundnut meal diet G1, G2, G3, G4, G5 and G6, were similarly supplemented with the same phytase levels used in experiment 1. Fish meal control (S0=G0) was not supplemented with phytase. Result showed that 250 FTU/g phytase showed the highest mean weight gain for both plants. In conclusion, the research has shown that the chemical nature of phytate, rather than its concentration and location, may influence the utilization of phosphorus for optimum growth in the fish by supplementing 250 FTU/g, with a range of available phosphorus requirement of between 0.75% (Y = 0.363 + 4.155X - 2.772X2, R2 = 0.759) and 0.80% (Y = 0.307 + 3.303X - 2.059X2, R2= 0.210)

Immobilised Phytase Production from Aspergillus foetidus MTCC 11682 Using an Optimized Media

Aim: Immobilised fungal phytase production from the novel strain Aspergillus foetidus MTTC 11682 and optimisation of cultural conditions for a better and continuous economic yield. Study Design: The study was designed based on the classical method of changing one independent variable while fixing all other at a certain level- one factor at a time, a close ended system for the optimisation of fermentation process. Methodology: Physical and nutritional parameters were optimised for phytase production and subjected to statistical analysis. Adsorption and Entrapment techniques were employed to immobilise the production strain. Results: The optimum physical conditions for augmenting the yield up to 6 days incubation period were as follows: pH of 3.5, 30ºC temperatures and 5% inoculum size. Amongst the nutritional parameters, lactose and sodium nitrate were found to be the best carbon and nitrogen sources. K++, Mg++, Mn++ and Fe++ ions supported the phytase production. TritonX 100 and tween 80 showed an inducing effect on the secretion of phytase enzyme. Immobilised fungal phytase production resulted in an increased yield of 32.5% with poly urethane foam (PUF) as the matrix. A scale up fermentation resulted in an activity of 52.7 FTU/mL for immobilised cells as compared to 25.5 FTU/mL by its free counterpart. Conclusion: Phytase produced in an optimised media employing immobilised Aspergillus foetidus 11682 on poly urethane foam cubes exhibited better phytase activity, improved stability and long shelf life.

Bacillus spp: una alternativa para la promoción vegetal por dos caminos enzimáticos / Bacillus spp: An alternative for plant promotion by two enzymatic pathways

Resumen Objetivo. Se realizó una revisión sobre las características de las fitasas y nitrogenasas de Bacillus spp. y sus opciones de uso como alternativa biofertilizante. El género Bacillus es secretor de proteínas y metabolitos eficientes para el control de plagas y enfermedades, promueve el crecimiento vegetal a través de la solubilización de fósforo y la producción de reguladores de crecimiento como el ácido indol acético; así mismo participa en la fijación de nitrógeno cuando hace parte de consorcios microbianos. Como biofertilizante es una opción amigable para el suelo y el ambiente que da respuesta a la necesidad de implementar la agricultura sostenible.

Abstract Objective. We conducted a review of the characteristics of the phytase and nitrogenase of Bacillus sp. and their potential use as an alternative biological fertilizer. The genus Bacillus is an efficient secretor of proteins and metabolites; to control pests and diseases, promote plant growth through the solubilization of phosphorus and production of growth regulators as acetic indole. Likewise it is involved in nitrogen fixation when it is part of microbial consortia. As biological fertilizer is a friendly option for the soil and the environment that responds to the need to implement sustainable agriculture.

Exogenous enzyme complex prevents intestinal soybean meal-induced enteritis in Mugil liza (Valenciennes, 1836) juvenile

ABSTRACT Four soybean meal-based diets containing increasing levels of an enzyme complex (E50, E100, E150 and E200 at 50, 100, 150 and 200 g ton-1, respectively) and one soybean meal-based diet without the enzyme complex (E0) were fed in triplicate to M. liza juveniles in a semi-static flow system with 20 fish per tank for 75 days. There were no differences between the treatments for animal performance parameters, but fish fed the enzyme complex treatment exhibited significantly (P<0.05) higher values of calcium bone retention compared with control fish. Although there was no relationship between bacterial counts in different sections of the gastrointestinal tract or enzyme levels, filamentous bacteria were increased in E50 compared with E150. All of the treatments resulted in higher bacterial counts in the stomach than in intestinal segments. Histological screening showed serious to moderate infiltration of inflammatory cells, modification in villus morphology and necrosis in some cases in fish fed the E0 diet. In addition, fish from the E0 treatment exhibited significantly (P<0.05) lower lipid deposition in the peritoneal cavity. Therefore, the use of low levels of exogenous enzyme is recommended in diets for M. liza when soybean meal is used as the main source of protein.

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.

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.

Endogenous losses and true digestibility of phosphorus in rice bran with or without phytase determined with piglets / Perdas endógenas e digestibilidade verdadeira do fósforo do farelo de arroz integral com ou sem fitase determinado com leitões

ABSTRACT: The aim of this study was to determine endogenous losses and true total digestibility of phosphorus (P) in piglets fed with diets containing rice bran (RB), and supplemented or not with phytase. The experiment was divided into three periods of 12 days each, seven for adaptation and five for total feces collection, with an interval of three days between periods. Twelve barrows with average body weights of 13.40±1.00kg at 46 days of age were distributed among six treatments consisting of diets formulated with three levels of RB (5%, 10%, and 20%) with or without the addition of 750FTUkg-1 of phytase. The amount of total daily feed was divided into four daily meals and water was available ad libitum. The method of total feces collection was used with ferric oxide as the fecal marker. The total consumption of P of RB origin was 0.37, 0.85 and 1.73gd-1 for concentrations of 5%, 10% and 20% of RB, respectively. Both RB and phytase levels had an effect on the apparent digestibility of P in RB. Results of the regression analysis of ingested P originating from the RB and absorbed P indicated endogenous losses of 527mg of Pkg-1 of dry matter intake, independent of phytase. Additionally, the true digestibility of P in RB was 47%, and it was not influenced by the inclusion of phytase. We concluded that the addition of 750FTU of phytase does not affect endogenous losses or the true digestibility of P in RB.

RESUMO: O objetivo do estudo foi determinar as perdas endógenas e a digestibilidade total verdadeira do fósforo (P) do farelo de arroz integral (FAI) com ou sem suplementação de fitase. O experimento foi dividido em três períodos de 12 dias cada, sendo, sete de adaptação e cinco de coleta total de fezes, com um intervalo de três dias entre os períodos. Utilizaram-se 12 suínos castrados com peso médio de 13,40±1,00kg e 46 dias de idade, distribuídos em seis tratamentos que consistiram de dietas formuladas com três níveis de FAI (5, 10 e 20%), com adição ou não de 750FTUkg-1 de fitase. A quantidade de ração total foi dividida em quatro refeições diárias e a água disponibilizada à vontade. Foi utilizado o método de coleta total de fezes, utilizando o óxido férrico como marcador fecal. O consumo de P com origem no FAI foi de 0,37, 0,85 e 1,73gd-1 para os teores de 5, 10 e 20% de FAI, respectivamente. Tanto os níveis FAI como a fitase exerceram efeitos na digestibilidade aparente do P do FAI. Os resultados da análise de regressão entre P ingerido com origem no FAI e P absorvido indicaram perdas endógenas de 527mg de Pkg-1 de matéria seca ingerida (DMI), independente do uso da fitase. Além disso, a digestibilidade verdadeira do P no FAI foi de 47% e não sofreu influência da inclusão de fitase. Conclui-se que a adição de 750FTU de fitase não influencia nas perdas endógenas e nem na digestibilidade verdadeira de P no FAI.

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.

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.

Characterization and expression of codon optimized soybean phytase gene in E. coli.

Phytic acid, the major storage form of phosphorus in plant seeds is degraded by the phytases to yield inositol and free phosphate, contributing thereby to the improved bioavailability of phytate phosphorus and essential minerals in plant foods and simultaneous reduction in phosphorus pollution of the terrestrial and aquatic ecosystems. As a possible strategy for altering seed phytate levels, the approach involving reduction of phytate content by ectopically expressing endogenous phytase gene during seed development of soybean (Glycine max L. cv. Pusa-20) was attempted in the present study. Semi-quantitative RT-PCR revealed the maximum expression of phytase gene transcripts in germinating cotyledons (~10 days after germinations), compared to other vegetative tissues. A full-length phytase cDNA was amplified from the germinating seedlings by splicing by overlap extension (SOE)-PCR and its sequence analysis revealed an open-reading-frame of 1644 bp, including an N terminal signal peptide of 28 amino acids. Predicted amino acid sequence (547-aa) of molecular mass 62 kDa on alignment with related purple acid phosphatases in other plants shared five conserved domains and seven invariant amino acids involved in coordination of the metals in the binuclear center of purple acid phosphatases. Owing to a large number of E. coli low-usage codons in soybean phytase gene, the modified gene was cloned into a prokaryotic expression vector pET-28a (+) and its expression in E. coli was confirmed by SDS-PAGE and Western blot analysis. Bioassay of the crude expression product in E. coli revealed a functional phytase gene, showing a great potential for developing low phytate transgenic soybean through its seed-specific overexpression in the early stages of seed development.

Isolation, Characterization of Phytase Producing Bacillus Subtils BtRS2 from the Rhizosphere Soil of Bt Cotton Field.

Soil samples of Bt Rhizosphere were collected from Bt cotton growing area of Andhra Pradesh, India and was used as a source material for isolation and screening of phytase producing bacteria. 19Bacteria were isolated from Bt Rhizosphere. Phytase enzyme activity of the cultures was screened on modified phytase solubulizing medium (MPSM). The result inferred that six isolates BtRS1 to BtRS6 were strongly positive in enzyme activity than six of other microorganisms while seven isolates were found negative and among all, Isolate BtRS2produced significantly higher phytase yield than other isolates and was chosen for species identification. Preliminary identification by microscopic and biochemical tests identified the isolate BtRS2 as Bacillus sp. Further, the identification of the strain was confirmed by subjecting it to 16S rRNA gene sequence analysis. The isolate BtRS2 showed high similarity to Bacillus subtilis sps and was thus designated as Bacillus subtilis BtRS2 The sequence is deposited under accession no. KC471313 in GeneBank.

Effects of Phytase Supplementation of Low Phosphorous Diets Included Olive Pulp and Date Pits on Productive Performance of Laying Hens, Egg Quality Traits and Some Blood Parameters.

Aims: This study was conducted to evaluate effects of phytase (E) supplementation of low phosphorous diets (NPP) included olive pulp (OP) and date pits (DP) on performance of laying hens, egg quality traits, blood parameters and excreta pH of laying hens. Study Design: Data were analyzed based on 2×2×2 factorial arrangements in completely randomized design using GLM procedure of SAS. Place and Duration of Study: The present experiment was done in Animal Science Department of Razi University, Kermanshah, Iran. All procedures used in this study were approved by the Animal Ethics Committee of Razi University and complied with the "Guidelines for the Care and Use of Animals in Research". Methodology: A total number of 288 Lohmann LSL-Lite laying hens was randomly divided in 48 cages (n=6). Based on a 2×2×2 factorial arrangement of treatments, 8 iso-caloric and iso-nitrogenous experimental diets (ME =2720 Kcal/Kg and CP=150 g/Kg) consisting of two levels of date pits and olive pulp (0, and 90 g/kg), two levels of dietary non-phytate phosphorus (NPP: 2.6 and 3.3 g/kg diet) and phytase (0 and 150 FTU/kg) were formulated. Hens in every randomly-selected six cages (replicates) allotted to feed on each of 8 experimental diets. Results: Dietary treatment did not have significant effect on egg production (EP) and feed intake (FI). Phytase numerically increased egg mass (EM) compared to control diets. Dietary inclusion of date pits and olive pulp significantly affected feed conversion ratio (FCR). Yolk index and Haugh unit were not significantly affected by dietary treatment (P>0.05). Diet inclusion of olive pulp and date pits numerically decreased eggshell weight and thickness in the first egg sampling (wk 3) in compared to control diet (P=.05). In the second egg sampling (wk 7), egg index, yolk index, Haugh unit, egg gravity and eggshell thickness were not significantly affected by dietary treatment. Dietary treatment did not have significant effect on blood parameters except for monocyte, so that a significant interaction between DO, P and E (p=0.01) was detected. Dietary treatment did not have significant effect on body weight changes (BWC) and excreta pH. Conclusion: From the results of the present study, it can be concluded that DP and OP can be included in diet of laying hens up to 9% with no adverse effect on birds' performance. However, diet inclusion of DP and OP has some adverse effects on egg yolk color and eggshell weight.

Efeito de estratégias nutricionais para redução de nutrientes poluidores nos dejetos sobre o desempenho e as características de carcaça de suínos / The effects of nutritional strategies to reduce nutrient pollution in manure on the performance and carcass traits of pigs

Avaliou-se o efeito de estratégias nutricionais sobre o desempenho e as características de carcaça de suínos dos 30 aos 100kg. Foram utilizados 120 suínos, distribuídos em delineamento experimental de blocos ao acaso, com cinco tratamentos e oito repetições de três animais por unidade experimental. Os tratamentos usados foram: dieta-controle = dieta com alta proteína bruta sem aminoácidos industriais; dieta baixa PB= dieta com baixa proteína bruta com aminoácidos industriais; dieta fitase = dieta-controle com suplementação de fitase; dieta orgânica = dieta-controle suplementada com minerais inorgânico-orgânicos; e dieta composta = dieta com baixa proteína bruta suplementada com aminoácidos industriais, fitase e minerais inorgânico-orgânicos. As dietas avaliadas não alteraram o desempenho e as características de carcaça dos suínos em comparação aos animais da dieta-controle. Conclui-se que a redução de proteína bruta, a suplementação com fitase e minerais inorgânico-orgânicos podem ser adotadas por não alterarem significativamente o desempenho nas fases de crescimento e terminação e as características de carcaças.

The objective was to evaluate the effect of different nutritional strategies on performance and carcass traits of pigs from 30 to 100kg. One hundred and twenty pigs were used, distributed in randomized blocks, with five treatments and eight replicates of three animals per experimental unit. Treatments were: control diet = corn-soybean based diet without synthetic amino acids; low crude protein diet = corn-soybean based diet with low protein and synthetic amino acids; phytase diet = control diet supplemented with phytase; organic diet = control diet supplemented with inorganic and organic minerals; and combination diet = diet with low protein supplemented with phytase and inorganic-organic minerals. Means were compared with the control diet by the Dunnett test. The diets evaluated did not alter the performance of pigs when compared to pigs fed the control diet. The reduction of crude protein, supplementation with phytase and inorganic-organic minerals can be adopted without significantly changing performance during the growing-finishing and carcass traits at 100kg.

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.

Fósforo fítico y actividad de fitasa en fórmulas infantiles basadas en cereales

El ácido fítico es un ácido orgánico presente en granos de cereales. El fósforo contenido en su molécula (PFi) no es absorbible por una baja solubilidad, aunque su biodisponibilidad puede incrementarse por la actividad de enzimas fitasas (AFi). Con el objetivo de cuantificar el contenido de PFi y AFi en suplementos elaborados a partir de cereales y destinados a infantes lactantes, se seleccionaron cinco fórmulas de amplia distribución en mercados locales,identificadas de acuerdo a los cereales base de su composición como: A (arroz), T (trigo), ATS (arroz, trigo y aislado de soya), ATM (arroz, trigo y maíz) y ATMS (arroz, trigo, maíz y aislado de soya). Se tomaron cinco muestras por fórmula, cada una correspondiente a un envase (400 a 500 g) elaborado en lotes diferentes y con fecha de vencimiento posterior al muestreo. La proteína cruda varió de 16,8 a 7,2%, con el mayor valor para ATS, y los menores para T y ATM (P< 0,01). El extracto etéreo mostró un rango de 0,31 a 0,75%, mientras la relación calcio:fósforo entre 1,6 en ATM y 1,1 para el resto. El PFi representó el 61,5% del fósforo total en T, con diferencias (P< 0,01) respecto al resto (39,9 ± 6,8%). Sólo se detectó AFi en ATM, T y ATS (570, 300 y 152 U/kg, respectivamente). Se concluye que la composición química se ajustó a lo reportado por los fabricantes, con un elevado contenido de PFi y una AFi fundamentalmente asociados a la presencia de trigo en las fórmulas.

Phytic phosphorus and phytase activity in cerealbased infant formulas. Phytic acid is an organic acid present in cereal grains. The phosphorus inside its molecule (PPhy) is not available because of its low solubility, though the bioavailability could be increased by the activity of phytase enzymes (PhyA). With the purpose of quantifying the PPhy and PhyA in supplements manufactured from cereals and intended for lactating infants, five formulas of wide distribution in local markets were selected and identified depending on the main vegetable ingredients as A (rice), T (wheat), ATS (rice, wheat and soy isolate), ATM (rice, wheat and maize) and ATMS (rice, wheat, maize and soy isolate). Five samples were taken from each formula, each one corresponded to a commercial brand (400 to 500 g), coming from different batches and before their expiration date. The crude protein ranged from 7.2 to 16.8%, with de highest value for ATS and the lowest for T and ATM (P< 0.01). Ether extract varied from 0.31 to 0.75%, while the calcium: phosphorus ratio from 1.6 for ATM, to 1.1 for the rest. The PPhy was 61.5% of the total phosphorus in T, with differences (P<0.01) compared to the other formulas (39.9 ± 6.8%).PhyA was only detected in ATS, T and ATM (152,300 and 570 U/kg, respectively). The chemical composition complied with the manufacturer report, with a high content of PPhy and PhyA associated to the wheat presence in the formula.

Enzimas exógenas em dietas de frangos de corte: desempenho / Exogenous enzymes in broilers fed diets: performance

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.

The experiment was conducted to evaluate the effect of enzyme efficiency in diets with and without nutrient reduction on the performance of broilers. Thousand and hundred forty male broiler chicks (Cobb 500®) were distributed in a completely randomized design with 4 treatments in a factorial 2x2 (two diets with different nutrient densities, with and without enzyme) with 8 replications of 45 birds. The diets were: positive control (normal) and a negative control (with a reduction of 4.3% and 4.5% of metabolizable energy, 16.7% and 17.7% of calcium and 35% and 42.7% of phosphorus, in initial and growth phases, respectively). The enzyme supplementation consisted of a combination of phytase enzyme (100g t-1) and amylase, xylanase and protease (500g t-1). It was evaluated the performance of birds in phases 1 to 21 and 1 to 42 days old. In the total period, the birds fed with negative control diet with added enzyme had feed intake (5.97%), body weight (8.47%), body weight gain (8.64%) and feed conversion (2.92%) higher (P<0.05) than compared to birds fed with negative control diet without enzyme supplementation. In addition, birds fed with control diet with added enzyme showed negative feed intake, average live weight and body weight gain similar (P>0.05) to the group fed with positive control diet with or without enzyme. It is concluded that the addition of exogenous enzymes in diets with reduced metabolizable energy, calcium and phosphorus, providing a feed intake, average live weight and weight gain similar to a diet with adequate levels for broiler.