Mannooligosaccharide production from açaí seeds by enzymatic hydrolysis: optimization through response surface methodology.

Recognized for its bioactive compounds, açaí has become a functional food, but it has a low pulp yield, and the seeds are the main waste. This study investigates the potential of açaí seeds (Euterpe oleracea Mart.) to produce mannooligosaccharides (MOS) through enzymatic hydrolysis. Using response surface methodology (RSM), the research optimizes MOS extraction while minimizing mannose production and reducing processing time, achieving MOS production of about 10 g/L, a value within the range of similar investigations. The RSM quadratic models establish correlations between MOS production (M2-M5) and enzymatic hydrolysis conditions, with R2 values ranging from 0.6136 to 0.9031. These models are used to emphasize MOS performance (M2-M5) while reducing mannose production, which also promotes profitability by reducing time. Experimental validation agrees with model predictions, highlighting optimal conditions near 40 °C, intermediate enzyme loading, and basic pH that effectively promotes MOS generation on mannose within an accelerated processing time frame. With predictions of experimental results within a margin of error of < 9%, the validity of the models was acceptable. This research contributes to the advancement of the understanding of the enzymatic hydrolysis of açaí seeds, which is a step toward the sustainable use of resources with a focus on process engineering aspects.

Unravelling biochemical and structural features of Bacillus licheniformis GH5 mannanase using site-directed mutagenesis and high-resolution protein crystallography studies.

Glycoside hydrolase family 5 (GH5) encompasses enzymes with several different activities, including endo-1,4-ß-mannosidases. These enzymes are involved in mannan degradation, and have a number of biotechnological applications, such as mannooligosaccharide prebiotics production, stain removal and dyes decolorization, to name a few. Despite the importance of GH5 enzymes, only a few members of subfamily 7 were structurally characterized. In the present work, biochemical and structural characterization of Bacillus licheniformis GH5 mannanase, BlMan5_7 were performed and the enzyme cleavage pattern was analyzed, showing that BlMan5_7 requires at least 5 occupied subsites to perform efficient hydrolysis. Additionally, crystallographic structure at 1.3 Å resolution was determined and mannoheptaose (M7) was docked into the active site to investigate the interactions between substrate and enzyme through molecular dynamic (MD) simulations, revealing the existence of a - 4 subsite, which might explain the generation of mannotetraose (M4) as an enzyme product. Biotechnological application of the enzyme in stain removal was investigated, demonstrating that BlMan5_7 addition to washing solution greatly improves mannan-based stain elimination.

Upgrading the Nutritional Value of PKC Using a Bacillus subtilis Derived Monocomponent ß-Mannanase.

Palm kernel cake (PKC) is an abundant side stream that can only be added to non-ruminant feed in small concentrations due to its content of antinutritional factors, mainly galactomannan, which cannot be digested by non-ruminants. ß-mannanases can be added to partially hydrolyze galactomannan to form mannose oligosaccharides, which are known to be prebiotic. We here investigate the action of a ß-mannanase from B. subtilis on PKC by colorimetry, NMR and fluorescence microscopy. The amount of mannan oligosaccharides in solution was significantly increased by the ß-mannanase and their degree of polymerization (DP) was significantly reduced. There was a dose-response behavior in that larger ß-mannanase concentrations increased the amount of soluble mannose oligosaccharides while reducing their average DP. Using confocal immunofluorescence microscopy, solubilization of galactomannan in PKC was clearly visualized. Images show a clear disruption of the cellulose and galactomannan structures of the PKC cell walls. We thus show in this study that using commercial dosages of ß-mannanase on PKC can lead to formation of prebiotic compounds. Thus, this study suggests that utilization of PKC in poultry feed formulation might be increased by addition of a ß-mannanase and would improve the return on investment.

Enzymatic preparation of manno-oligosaccharides from locust bean gum and palm kernel cake, and investigations into its prebiotic activity

BACKGROUND: Manno-oligosaccharides (MOS) is known as a kind of prebiotics. Mannanase plays a key role for the degradation of mannan to produce MOS. In this study, the mannanases of glycoside hydrolase (GH) families 5 Man5HJ14 and GH26 ManAJB13 were employed to prepare MOS from locust bean gum (LBG) and palm kernel cake (PKC). The prebiotic activity and utilization of MOS were assessed in vitro using the probiotic Lactobacillus plantarum strain. RESULTS: Galactomannan from LBG was converted to MOS ranging in size from mannose up to mannoheptose by Man5HJ14 and ManAJB13. Mannoheptose was got from the hydrolysates produced by Man5HJ14, which mannohexaose was obtained from LBG hydrolyzed by ManAJB13. However, the same components of MOS ranging in size from mannose up to mannotetrose were observed between PKC hydrolyzed by the mannanases mentioned above. MOS stability was not affected by high-temperature and high-pressure condition at their natural pH. Based on in vitro growth study, all MOS from LBG and PKC was effective in promoting the growth of L. plantarum CICC 24202, with the strain preferring to use mannose to mannotriose, rather than above mannotetrose. CONCLUSIONS: The effect of mannanases and mannan difference on MOS composition was studied. All of MOS hydrolysates showed the stability in adversity condition and prebiotic activity of L. plantarum, which would have potential application in the biotechnological applications.

Reserve mobilization dynamics and degradation pattern of mannan-rich cell walls in the recalcitrant seed of Mauritia flexuosa (Arecaceae).

Most seeds store reserves, which mobilization after germination is complex and diversified among plant species. Information on the reserve mobilization in recalcitrant seeds (i.e., intolerant to desiccation) is scarce. The aim of this work was to characterize the dynamics of reserve mobilization and the degradation pattern of the endospermic cell walls in the recalcitrant seeds of the neotropical palm Mauritia flexuosa. Biometric, anatomical, histochemical, ultrastructural and immunocytochemistry assessments were performed in the endosperm and haustorium (structure of the seedling involved in reserve mobilization), during germination and throughout seedling development. Endo-ß-mannanase activity was assessed. The main reserves stored in the seeds are mucilage in the living protoplast and, mainly, heteromannans in the thick cell walls of the endosperm cells. The reserve mobilization extends for about 180 days, in four phases. During germination, the embryonic reserves are catabolized, which induces the mobilization of the endosperm by establishing the flow of water and carrying substances to the haustorium. After germination, the cells of the endosperm actively control the process of their degradation, which results in the formation of the digestion zone. The growth of the haustorium promotes the crushing of endospermic cells and facilitates the entry of substances via the apoplastic route. The pattern of degradation of endospermic cells involves three phases: 1) mobilization of the vacuolar content by symplastic route; 2) increased vacuole turgor pressure, directing the content of the cytoplasm to the cell walls; 3) degradation of cell walls.

Effects of -mannanase on Egg Production Performance, Egg Quality, Intestinal Microbiota, Viscosity, and Ammonia Concentration in Laying Hens

Two experiments were conducted to evaluate the effects of -mannanase on egg production performance, egg quality, intestinal microbiota, viscosity, and ammonia concentration in laying hens. In Exp. 1, two hundred and seventy 30-wk-old Hy-Line Brown laying hens were assigned to 6 diets arranged in a 3 × 2 factorial of three levels of MEn and CP [(a corn-soybean meal based diet (HEHP), a diet containing 50 kcal of MEn/kg and 1.0% less energy and CP than the HEHP (MEMP), and a diet containing 80 kcal of MEn/kg and 1.5% less energy and CP than HEHP (LELP)], and -mannanase supplementation (0 or 0.04%). In Exp. 2, A total of two hundred and sixteen62-wk-old Hy-Line Brown laying hens were assigned to 6 dietary treatments in a 3 × 2 factorial arrangement. In Exp. 1, -mannanase supplementation increased egg production rate in hens fed LELP diet, but not in those fed HEHP or MEMP diet (interaction, p 0.01), and the interaction was significant (p 0.01) for egg mass. -mannanase supplementation decreased (p 0.05) ammonia concentration. In Exp. 2, the supplementation of -mannanase increased egg production rate and egg mass in hens fed LELP diet, whereas no differences were found in those fed HEHP or MELP diet (interaction, p 0.01). The supplementation of -mannanase in a lower energy and protein diet resulted in similar production performance when compared to high-energy and high-protein diet during early and late stages of egg production.(AU)

Effects of -mannanase on Egg Production Performance, Egg Quality, Intestinal Microbiota, Viscosity, and Ammonia Concentration in Laying Hens

Two experiments were conducted to evaluate the effects of -mannanase on egg production performance, egg quality, intestinal microbiota, viscosity, and ammonia concentration in laying hens. In Exp. 1, two hundred and seventy 30-wk-old Hy-Line Brown laying hens were assigned to 6 diets arranged in a 3 × 2 factorial of three levels of MEn and CP [(a corn-soybean meal based diet (HEHP), a diet containing 50 kcal of MEn/kg and 1.0% less energy and CP than the HEHP (MEMP), and a diet containing 80 kcal of MEn/kg and 1.5% less energy and CP than HEHP (LELP)], and -mannanase supplementation (0 or 0.04%). In Exp. 2, A total of two hundred and sixteen62-wk-old Hy-Line Brown laying hens were assigned to 6 dietary treatments in a 3 × 2 factorial arrangement. In Exp. 1, -mannanase supplementation increased egg production rate in hens fed LELP diet, but not in those fed HEHP or MEMP diet (interaction, p 0.01), and the interaction was significant (p 0.01) for egg mass. -mannanase supplementation decreased (p 0.05) ammonia concentration. In Exp. 2, the supplementation of -mannanase increased egg production rate and egg mass in hens fed LELP diet, whereas no differences were found in those fed HEHP or MELP diet (interaction, p 0.01). The supplementation of -mannanase in a lower energy and protein diet resulted in similar production performance when compared to high-energy and high-protein diet during early and late stages of egg production.

Dietary β-mannanase decreases cloacal temperature of broiler chickens under hot conditions without affecting growth performance / El consumo de β-mananasa disminuye la temperatura cloacal de pollos de engorde bajo condiciones de calor sin afectar su rendimiento de crecimiento / O consumo de β-mananase diminui a temperatura da cloaca do frango de corte em condições de calor sem afetar seu desempenho do crescimento

Abstract Background: High amounts of nonstarch polysaccharides in the diet may increase the amounts of fermentative materials in the hindgut, leading to an increase in fermentative heat production. Dietary β-mannanase is reported to decrease antinutritional effects of β-mannans, such as the potential increase of body heat; however, its efficacy on broiler chickens raised under hot climatic conditions has not been investigated. Objective: To investigate the effects of dietary β-mannanase on growth performance, cloacal temperature, relative lymphoid organ weight, and blood characteristics of broiler chickens raised under hot climatic conditions. Methods: A total of 1,701 1-day-old Ross 308 broiler chickens were randomly allotted to one of three dietary treatments with nine replicates. A basal diet was prepared and added with β-mannanase at 0.05 or 0.10% inclusion levels. The experiment was conducted for 30 days. Average room temperature was 28.8 ± 1.74 ˚C and average relative humidity (RH) was 76.1 ± 11.49% during the experiment. Results: Growth performance of broiler chickens raised under hot climatic conditions was not affected by β-mannanase inclusion. Cloacal temperature decreased at the end of experiment (linear, p<0.05) with increasing inclusion levels of dietary β-mannanase. Increasing inclusion levels of β-mannanase tended to increase (linear, p=0.076) the relative weight of thymus, but had no effects on the relative weight of spleen and bursa of Fabricius. Blood characteristics were not influenced by dietary β-mannanase. Conclusion: Increasing inclusion levels of β-mannanase decrease cloacal temperature; however, it does not directly influence growth performance nor alleviates the heat stress of broiler chickens raised under hot climatic conditions.

Resumen Antecedentes: Altas cantidades de carbohidratos no almidonosos en la dieta pueden aumentar la cantidad de materiales fermentativos en el intestino posterior, aumentando la producción de calor fermentativo. La β-mananasa dietaria disminuye los efectos antinutricionales de los β-mananos, tales como el potencial incremento de la producción calorica; Sin embargo, no se ha investigado su eficacia en pollos de engorde criados bajo condiciones de calor ambiental. Objetivo: Investigar los efectos de la β-mananasa en la dieta sobre el crecimiento, la temperatura cloacal, el peso relativo de los órganos linfoides y las características sanguíneas de pollos de engorde criados bajo condiciones climáticas calientes. Métodos: Un total de 1.701 pollos de engorde de 1 día de edad (Ross 308) fueron asignados al azar a uno de tres tratamientos dietarios con nueve repeticiones. A una dieta basal se le adicionó β-mananasa en niveles de inclusión de 0,05 o 0,10%. El experimento duró 30 días. La temperatura ambiente durante el experimento fue de 28,8 ± 1,74 ˚C y la humedad relativa de 76,1 ± 11,49%. Resultados: La inclusión de β-mananasa no afecto el rendimiento de los pollos. La temperatura cloacal medida al final del experimento disminuyó (lineal, p<0,05) con niveles de inclusión dietarios crecientes de β-mananasa. Niveles incrementales de β-mananasa tendieron a aumentar (lineal, p=0,076) el peso relativo del timo, pero no hubo efecto sobre el peso del bazo o la bursa de Fabricio. La β-mananasa no influenció las características de la sangre. Conclusión: Niveles incrementales de β-mananasa disminuyen la temperatura cloacal, aunque afectan el crecimiento ni alivian el estrés térmico del pollo de engorde criado bajo condiciones climáticas calientes.

Resumo Antecedentes: Altas quantidades de polissacáridos não amiláceos na dieta podem aumentar as quantidades de materiais fermentativos no intestino posterior, levando a um aumento na produção de calor fermentativo. A β-mananase dietética é relatada para diminuir os efeitos antinutricionais da β-manana, tal como o possível aumento da produção de calor; Entretanto, não tem sido investigada a eficácia de la em frangos de corte criados em condições climáticas quentes. Objetivo: Investigar os efeitos da β-mananase na dieta sobre o desempenho do crescimento, a temperatura cloacal, o peso relativo dos órgãos linfóides e as características de sanguíneas de frangos de corte criados em condições climáticas quentes. Métodos: Um total de 1.701 frangos de corte de um dia de idade (Ross 308) foram distribuídos aleatoriamente em um dos três tratamentos dietéticos com nove repetições. A dieta basal foi preparada e a β-mananase foi adicionada à dieta basal com níveis de inclusão de 0,05 ou 0,10%. O experimento foi conduzido durante 30 dias. A temperatura ambiente média foi de 28,8 temperatura ahumidade relativa (HR) média foi de 76,1 ± 11,49% durante o experimento. Resultados: O desempenho de crescimento dos frangos de corte criados em condições climáticas quentes não foi afetado pelos níveis de inclusão de β-mananase. A temperatura cloacal medida no final do experimento foi diminuída (linear, p<0,05) com níveis crescentes de inclusão de β-mananase na dieta. O aumento dos níveis de inclusão da β-mananase tendeu a aumentar (linear, p=0,076) o peso relativo do timo, mas não teve efeitos sobre o peso relativo do baço e da bursa de Fabricius. As características do sangue não foram influenciadas pela β-mananase dietética. Conclusão: Os níveis crescentes de inclusão de β-mananase diminuem a temperatura cloacal; entretanto, isso não influencia diretamente o desempenho do crescimento e alivia o estresse térmico dos frangos de corte criados em condições climáticas quentes.

The Role of ß-Mannanase (Hemicell) in Improving Poultry Productivity, Health and Environment

The poultry nutritionists constantly attempt to identify and alleviate the factors responsible for adverse effects on nutrient utilization and production performance in avian species, which are necessary for successful productivity of poultry. Enhanced feed utilization can reduce levels of some nutrients in the diet with concomitant mitigation in nutrient excretion into the environment, reduction of eutrophication and acidification potentials of excreta. Commercial enzymes have been used to improve feed efficiency and utilization in poultry. Among many anti-nutritional factors, the existence of ß-mannans in poultry feed ingredients including soybean and other leguminous seeds has been associated with negative effects on nutrient digestibility and high intestinal viscosity that adversely affects innate immunity and microbial proliferation in poultry gut. The ß-mannanase (a commercial product named as Hemicell) can hydrolyze ß-mannan, an anti-nutritional fiber present in many poultry feed ingredients. Supplement of ß-mannanase to ß-mannan-rich diets may boost the population of intestinal beneficial bacteria, increase the digestibility of mannans, enhance the immunity, suppresses the growth of harmful intestinal bacteria, enhance the digestion and absorption of nutrients in intestinal tracts and reduce the environmental pollution due to poultry excreta. Supplementation of -mannanase at the level of 200 and 400 mg/kg in poultry diets has positively improved blood glucose and anabolic hormone homeostasis, FCR, digestible energy, and digestible amino acids. This review describes the promising beneficial effects of -mannanase, which may be used in the poultry feed industry for economic benefits. Another objective of this review is to explore the underlying mechanisms of ß-mannanase that can influence growth, digestion coefficients of nutrients, health and metabolism of nutrients in poultry birds and also the knowledge regarding the useful application of this feed enzyme in the commercial poultry feed industry.

The Role of ß-Mannanase (Hemicell) in Improving Poultry Productivity, Health and Environment

The poultry nutritionists constantly attempt to identify and alleviate the factors responsible for adverse effects on nutrient utilization and production performance in avian species, which are necessary for successful productivity of poultry. Enhanced feed utilization can reduce levels of some nutrients in the diet with concomitant mitigation in nutrient excretion into the environment, reduction of eutrophication and acidification potentials of excreta. Commercial enzymes have been used to improve feed efficiency and utilization in poultry. Among many anti-nutritional factors, the existence of ß-mannans in poultry feed ingredients including soybean and other leguminous seeds has been associated with negative effects on nutrient digestibility and high intestinal viscosity that adversely affects innate immunity and microbial proliferation in poultry gut. The ß-mannanase (a commercial product named as Hemicell) can hydrolyze ß-mannan, an anti-nutritional fiber present in many poultry feed ingredients. Supplement of ß-mannanase to ß-mannan-rich diets may boost the population of intestinal beneficial bacteria, increase the digestibility of mannans, enhance the immunity, suppresses the growth of harmful intestinal bacteria, enhance the digestion and absorption of nutrients in intestinal tracts and reduce the environmental pollution due to poultry excreta. Supplementation of -mannanase at the level of 200 and 400 mg/kg in poultry diets has positively improved blood glucose and anabolic hormone homeostasis, FCR, digestible energy, and digestible amino acids. This review describes the promising beneficial effects of -mannanase, which may be used in the poultry feed industry for economic benefits. Another objective of this review is to explore the underlying mechanisms of ß-mannanase that can influence growth, digestion coefficients of nutrients, health and metabolism of nutrients in poultry birds and also the knowledge regarding the useful application of this feed enzyme in the commercial poultry feed industry.(AU)

Enzyme complex and Saccharomyces cerevisiae in diets for broilers in the initial phase / Complexo enzimático e “Saccharomyces cerevisiae ” em dietas para frangos de corte na fase inicial

This study aimed at evaluating the use of exogenous enzymes in diets with Saccharomyces cerevisiae and their impact on zootechnical performance, carcass yield, intestinal histomorphometry and of broiler diets in the initial phase. A completely randomized design was used in a 2×3 + 1 factorial arrangement, with two levels of enzyme complex (EC), (0 and 200g / ton), three yeast levels (0, 6 and 12%) and a control diet, making up seven treatments, with five replicates of 20 broilers per experimental unit. We evaluated the performance (feed intake, weight gain and feed conversion ratio), carcass yield and cuts, histomorphometry of the small intestine (height, circumference and width of villi, height and width of the crypt, thickness of the intestinal muscle wall and villi/crypt relationship). From 1 to 7 and 1 to 21 days, the inclusion of yeast led to reduced broiler performance. At 21 days, the addition of EC resulted in an increase of (p 0.05) in the thickness of the muscular wall of the duodenum and decreased the width of the crypt in the ileum. The 12% level of yeast without the EC provided a thicker jejunum intestinal muscle wall when compared to the positive control. There was no significant effect on carcass yield and cuts between treatments. In conclusion, the inclusion of yeast reduces performance from 1-21 days. The enzyme complex and yeast does not change the performance or carcass yield, however, it does bring benefits to the intestinal mucosa.

Objetivou-se avaliar o uso de enzimas exógenas em dietas com Saccharomyces cerevisiae sobre o desempenho zootécnico, rendimento de carcaça e histomorfometria intestinal de frangos de corte na fase inicial. Utilizou-se o delineamento inteiramente casualizado em esquema fatorial 2×3+1, sendo dois níveis de complexo enzimático (CE), (0 e 200g/ton), três níveis de levedura (0, 6 e 12%) e uma dieta controle, perfazendo sete tratamentos, cinco repetições de 20 aves por unidade experimental. Foram avaliados o desempenho (consumo de ração, ganho de peso e conversão alimentar), rendimento de carcaça e cortes e a histomorfometria do intestino delgado (altura, perímetro e largura de vilo, altura e largura de cripta, espessura da parede muscular intestinal e relação vilo/cripta). Na fase de 1 a 7 e de 1 a 21 dias, a inclusão de levedura na dieta promoveu redução no desempenho dos frangos. Aos 21 dias a adição de CE resultou em aumento (p 0,05) da espessura da parede muscular do duodeno, e reduziu a largura da cripta no íleo. O nível de 12% de levedura sem o CE proporcionou parede muscular intestinal do jejuno mais espessa quando comparada ao controle positivo. Não houve efeito significativo para rendimento de carcaça e cortes entre os tratamentos. Conclui-se que, a inclusão de levedura reduz o desempenho de 1 a 21 dias. A adição de complexo enzimático e levedura em dietas para frangos de corte não melhora o desempenho e rendimento de carcaça, todavia, beneficia à mucosa intestinal.

Enzyme complex and Saccharomyces cerevisiae in diets for broilers in the initial phase / Complexo enzimático e “Saccharomyces cerevisiae ” em dietas para frangos de corte na fase inicial

This study aimed at evaluating the use of exogenous enzymes in diets with Saccharomyces cerevisiae and their impact on zootechnical performance, carcass yield, intestinal histomorphometry and of broiler diets in the initial phase. A completely randomized design was used in a 2×3 + 1 factorial arrangement, with two levels of enzyme complex (EC), (0 and 200g / ton), three yeast levels (0, 6 and 12%) and a control diet, making up seven treatments, with five replicates of 20 broilers per experimental unit. We evaluated the performance (feed intake, weight gain and feed conversion ratio), carcass yield and cuts, histomorphometry of the small intestine (height, circumference and width of villi, height and width of the crypt, thickness of the intestinal muscle wall and villi/crypt relationship). From 1 to 7 and 1 to 21 days, the inclusion of yeast led to reduced broiler performance. At 21 days, the addition of EC resulted in an increase of (p 0.05) in the thickness of the muscular wall of the duodenum and decreased the width of the crypt in the ileum. The 12% level of yeast without the EC provided a thicker jejunum intestinal muscle wall when compared to the positive control. There was no significant effect on carcass yield and cuts between treatments. In conclusion, the inclusion of yeast reduces performance from 1-21 days. The enzyme complex and yeast does not change the performance or carcass yield, however, it does bring benefits to the intestinal mucosa.(AU)

Objetivou-se avaliar o uso de enzimas exógenas em dietas com Saccharomyces cerevisiae sobre o desempenho zootécnico, rendimento de carcaça e histomorfometria intestinal de frangos de corte na fase inicial. Utilizou-se o delineamento inteiramente casualizado em esquema fatorial 2×3+1, sendo dois níveis de complexo enzimático (CE), (0 e 200g/ton), três níveis de levedura (0, 6 e 12%) e uma dieta controle, perfazendo sete tratamentos, cinco repetições de 20 aves por unidade experimental. Foram avaliados o desempenho (consumo de ração, ganho de peso e conversão alimentar), rendimento de carcaça e cortes e a histomorfometria do intestino delgado (altura, perímetro e largura de vilo, altura e largura de cripta, espessura da parede muscular intestinal e relação vilo/cripta). Na fase de 1 a 7 e de 1 a 21 dias, a inclusão de levedura na dieta promoveu redução no desempenho dos frangos. Aos 21 dias a adição de CE resultou em aumento (p 0,05) da espessura da parede muscular do duodeno, e reduziu a largura da cripta no íleo. O nível de 12% de levedura sem o CE proporcionou parede muscular intestinal do jejuno mais espessa quando comparada ao controle positivo. Não houve efeito significativo para rendimento de carcaça e cortes entre os tratamentos. Conclui-se que, a inclusão de levedura reduz o desempenho de 1 a 21 dias. A adição de complexo enzimático e levedura em dietas para frangos de corte não melhora o desempenho e rendimento de carcaça, todavia, beneficia à mucosa intestinal.(AU)

Hormonal profile and the role of cell expansion in the germination control of Cerrado biome palm seeds.

Little information is currently available concerning the mechanisms controlling palm seed germination. We compared the anatomical and physiological aspects of seeds of two neotropical palm species showing different levels of dormancy. The seeds of Attalea vitrivir and Butia capitata were evaluated for the endogenous contents of hormones (ABA, GAs, CKs, BRs, IAA, JA, SA and the ethylene precursor ACC) in their cotyledonary petiole and operculum (structures involved in germination control), the force necessary to displace the operculum, endo-ß-mannanase activities, and embryo cell elongation. The analyses were carried out on with intact dry and imbibed seeds as well as with seeds with the operculum mechanically removed, 2, 5 and 10 days after sowing. The germinabilities of the intact seeds of A. vitrivir and B. capitata were 68% and 3%, respectively; the removal of the operculum increased germination to more than 90% in both species. Reductions of ABA and increases in GAs contents coincided with cell elongation, although there is no evidence that hormonal balance and endo-ß-mannanase activity are involved in operculum weakening. The ratio between the embryo length and the force required for operculum displacement (EL/OF) was found to be 1.9 times greater in A. vitrivir than in B. capitata, which means that very small elongations in each cell would be sufficient to promote germination, resulting in a lower level of dormancy in the former species. EL/OF and cell growth control are therefore important for defining dormancy level in palm seeds.

Roles of the haustorium and endosperm during the development of seedlings of Acrocomia aculeata (Arecaceae): dynamics of reserve mobilization and accumulation.

The mobilization of palm seed reserves is a complex process because of the abundance and diversity of stored compounds and results from the development of a highly specialized haustorium. This work focused on the important Neotropical oleaginous palm Acrocomia aculeata, with the aim of defining phases of seedling development associated with mobilization of reserves and elucidating the role of haustorium and endosperm in this process. Standard methods were performed, including biometric, anatomical, and histochemical analyses, as well as the evaluation of the activities of the enzymes endo-ß-mannanase and lipase, throughout the reserve mobilization in seeds during germination and in seedlings. Seeds of A. aculeata stored large quantities of proteins, lipids, and polysaccharides in the embryo and endosperm. The mobilization of reserves initiated in the haustorium during germination and subsequently occurred in the endosperm adjacent to the haustorium, forming a gradually increasing zone of digestion. Proteins and polysaccharides were the first to be mobilized, followed by lipids and cell wall constituents. The haustorium activates and controls the mobilization, forming transitory reserves and translocating them to the vegetative axis, while the endosperm, which also has an active role, serves as a site of intense enzymatic activity associated with protein bodies. Seedling development can be described as occurring in six phases over a long period (approximately 150 days) due to the large amount of seed reserves. This process exhibits an alternation between stages of accumulation and translocation of protein, lipid, and carbohydrate reserves in the haustorium, which favors the seedling establishment and the reproductive success of the species.

Effect of the addition of ß-mannanase on the performance, metabolizable energy, amino acid digestibility coefficients, and immune functions of broilers fed different nutritional levels.

Three experiments were conducted to evaluate the effect of ß-mannanase BM: supplementation on the performance, metabolizable energy, amino acid digestibility, and immune function of broilers. A total of 1,600 broilers were randomly distributed in a 4 × 2 factorial arrangement (4 nutritional levels × 0 or 500 g/ton BM), with 10 replicates and 20 broilers per pen. The same design was used in the energy and digestibility experiments with 8 and 6 replicates, respectively, and 6 broilers per pen. The nutritional levels : NL : were formulated to meet the nutritional requirements of broilers : NL1 : ; reductions of 100 kcal metabolizable energy : NL2 : ; 3% of the total amino acids (NL3); and 100 kcal metabolizable energy and 3% total amino acids (NL4) from NL1. The serum immunoglobulin (Ig) concentration was determined in two broilers per pen, and these broilers were slaughtered to determine the relative weight of spleen, thymus, and bursa of Fabricius. Throughout the experiment, the lower nutritional levels reduced (P < 0.05) body weight gain : BWG : and increased (P < 0.05) feed conversion : FCR : for the NL4 treatment. The BM increased (P < 0.05) the BWG values and improved (P < 0.05) the FCR of the broilers. The apparent metabolizable energy corrected for nitrogen balance (AMEn) values were reduced (P < 0.05) for NL2 and NL3. The BM increased (P < 0.05) the AMEn values and reduced (P < 0.05) the excreted nitrogen. NL3 and NL4 reduced (P < 0.05) the true ileal digestibility coefficients (TIDc) of the amino acids cystine and glycine, and BM increased (P < 0.05) the TIDc for all amino acids. The addition of BM reduced (P < 0.05) the relative weights of the spleen and bursa. NL2 increased (P < 0.05) the Ig values, whereas BM reduced (P < 0.05) the serum IgA, IgG, and IgM values of the broilers. This study indicates that using suboptimal nutrient levels leads to losses in production parameters, whereas BM-supplemented diets were effective in improving performance, energy values, and TIDc levels of amino acids and immune response of broilers.

Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest

ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE) and one bacterium (I4) strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes.

Effect of hydrolyzed copra meal separately or in combination with Bacillus cereus var. toyoi on growth performance of broiler chickens / Efeito da farinha hidrolisada da torta de coco, isoladamente ou em combinação com Bacillus cereus var. toyoi, sobre o desempenho de frangos de corte

The effects of mannanase-hydrolyzed copra meal (MCM) and MCM + probiotic Bacillus cereus var. toyoi (TY) on growth performance and gut morphometry of broiler chickens were investigated. A total of 1120-one-day-old Cobb chicks were distributed in a completely randomized design with 4 diet treatment groups. Dietary treatments were (1) negative control; (2) positive control (avilamycin 10 ppm); (3) 0.1% MCM for basal diets (4) 0.1% MCM + 0.05% TY. The best feed conversion ratio (FCR), body weight (BW), productivity index (PI) was obtained with 0.1% MCM + 0.05% TY at 42 days of age. With regard to productivity index, every supplementation group had a better rate when compared to that in negative group. Although 0.1% MCM supplemented alone is worse than positive group, it reveals a significantly better value than when it is combined with 0.1% MCM and 0.05% TY. The combination of mannanase-hydrolyzed copra meal with Bacillus cereus var. toyoi improved broiler performance and duodenum and jejunum mucous morphometry, when compared to the negative, positive and 0.1% MCM alone supplementation groups. The combination of MCM and TY probiotics is capable of improving intestinal morphology by behaving like a good growth promoter, with possibility of being an alternative to antibiotics.(AU)

Foram investigados os efeitos da farinha hidrolisada da torta de coco (MCM) e MCM + probiótico Bacillus cereus var. toyoi (TY) sobre o desempenho e morfometria intestinal de frangos de corte . Foram utilizados 1.120 pintos de corte da linhagem Cobb 500, distribuídos em delineamento inteiramente casualizado, com quatro tratamentos e dez repetições de 28 aves cada. Os tratamentos foram: (1) controle negativo, (2) controle positivo (avilamicina 10 ppm), (3) 0,1% MCM na dieta controle negativo, (4) 0,1% MCM + 0,05% TY na dieta controle negativo. A melhor conversão alimentar (CA), o peso corporal (PC) e o índice de produtividade (IP) foram obtidos no tratamento com 0,1% MCM + 0,05% TY aos 42 dias de idade. A combinação da farinha hidrolisada da torta de coco com o Bacillus cereus var. toyoi melhorou o desempenho e a morfometria da mucosa do duodeno e do jejuno, quando comparado com os grupos controle negativo, positivo e 0,1 % MCM isoladamente. A combinação do MCM com o TY tem potencial para melhorar a morfologia intestinal, comportando-se como um bom promotor de crescimento com possibilidade de alternativa aos antibióticos.(AU)

Effect of hydrolyzed copra meal separately or in combination with Bacillus cereus var. toyoi on growth performance of broiler chickens / Efeito da farinha hidrolisada da torta de coco, isoladamente ou em combinação com Bacillus cereus var. toyoi, sobre o desempenho de frangos de corte

The effects of mannanase-hydrolyzed copra meal (MCM) and MCM + probiotic Bacillus cereus var. toyoi (TY) on growth performance and gut morphometry of broiler chickens were investigated. A total of 1120-one-day-old Cobb chicks were distributed in a completely randomized design with 4 diet treatment groups. Dietary treatments were (1) negative control; (2) positive control (avilamycin 10 ppm); (3) 0.1% MCM for basal diets (4) 0.1% MCM + 0.05% TY. The best feed conversion ratio (FCR), body weight (BW), productivity index (PI) was obtained with 0.1% MCM + 0.05% TY at 42 days of age. With regard to productivity index, every supplementation group had a better rate when compared to that in negative group. Although 0.1% MCM supplemented alone is worse than positive group, it reveals a significantly better value than when it is combined with 0.1% MCM and 0.05% TY. The combination of mannanase-hydrolyzed copra meal with Bacillus cereus var. toyoi improved broiler performance and duodenum and jejunum mucous morphometry, when compared to the negative, positive and 0.1% MCM alone supplementation groups. The combination of MCM and TY probiotics is capable of improving intestinal morphology by behaving like a good growth promoter, with possibility of being an alternative to antibiotics.

Foram investigados os efeitos da farinha hidrolisada da torta de coco (MCM) e MCM + probiótico Bacillus cereus var. toyoi (TY) sobre o desempenho e morfometria intestinal de frangos de corte . Foram utilizados 1.120 pintos de corte da linhagem Cobb 500, distribuídos em delineamento inteiramente casualizado, com quatro tratamentos e dez repetições de 28 aves cada. Os tratamentos foram: (1) controle negativo, (2) controle positivo (avilamicina 10 ppm), (3) 0,1% MCM na dieta controle negativo, (4) 0,1% MCM + 0,05% TY na dieta controle negativo. A melhor conversão alimentar (CA), o peso corporal (PC) e o índice de produtividade (IP) foram obtidos no tratamento com 0,1% MCM + 0,05% TY aos 42 dias de idade. A combinação da farinha hidrolisada da torta de coco com o Bacillus cereus var. toyoi melhorou o desempenho e a morfometria da mucosa do duodeno e do jejuno, quando comparado com os grupos controle negativo, positivo e 0,1 % MCM isoladamente. A combinação do MCM com o TY tem potencial para melhorar a morfologia intestinal, comportando-se como um bom promotor de crescimento com possibilidade de alternativa aos antibióticos.

Armazenamento e qualidade de sementes de tomate enriquecidas com micronutrientes e reguladores de crescimento / Storage and quality of tomato seeds enriched with micronutrients and growth regulators

O estudo foi desenvolvido na Universidade Federal de Lavras, em Lavras, MG, com o objetivo de avaliar o efeito da aplicação de micronutrientes e reguladores de crescimento na germinação, no vigor, na atividade de algumas enzimas e no teor de proteínas totais em sementes de tomate, durante o armazenamento. As sementes foram tratadas com os produtos Starter®, Cellerate® e Stimulate® nas dosagens correspondentes a 0 por cento, 50 por cento, 100 por cento, 150 por cento e 200 por cento da dose recomendada pelo fabricante, utilizando a técnica de peliculização. As avaliações foram realizadas aos zero, 6 e 12 meses de armazenamento, pelos seguintes parâmetros: porcentagem de germinação, porcentagem e índice de velocidade de emergência, atividade das enzimas endo-â-mananase e esterase, teor de proteínas totais e sanidade. Foi utilizado o delineamento inteiramente casualizado, em esquema fatorial 3 x 5 x 3 ( 3 produtos x 5 doses x 3 épocas de armazenamento), com quatro repetições de 50 sementes por tratamento. Concluiu-se que a combinação dos reguladores de crescimento contidos no produto Stimulate® promove aumento na velocidade de emergência das plântulas de tomate quando aplicados na dose recomendada e na pré-semeadura; a atividade da enzima esterase aumenta com o período de armazenamento das sementes de tomate, indicando aumento no processo de deterioração; o revestimento enriquecido com os micronutrientes e reguladores de crescimento contidos nos produtos Starter®, Cellerate® e Stimulate® e o armazenamento interferem na atividade da enzima endo-â-mananase em sementes de tomate; há um aumento no teor de proteínas totais em sementes de tomate com o aumento do período de armazenamento.

The objective of this study was to evaluate the effect of the application of micronutrients and growth regulators on the germination, vigor, activity of some enzymes, and on the total protein contents in tomato seeds during the storage. The seeds were treated with the chemicals Starter®, Cellerate®, and Stimulate® at the dosages corresponding to 0 percent, 50 percent, 100 percent, 150 percent, and 200 percent of the dose recommended by the manufacturer, utilizing the film-coating technique. The evaluations were performed at 0, 6, and 12 months of storage by the following parameters: percentage of germination, percentage and emergency velocity rate, activity of enzymes endo-â-mannanase and esterase, total protein content, and health. The completely randomized design in a 3 x 5 x 3 factorial structure was utilized, namely, three chemicals, five doses, and three storage periods with four replicates of 50 seeds per treatment. Coating with the chemicals based on micronutrients and growth regulators and storage interfere on the activity of enzyme endo-â-mannanase in the tomatoes and seeds; the activity of enzyme esterase increases with the storage of tomato seeds, pointing to an increase in the deterioration process; there is an increase in the total protein content in tomato seeds throughout the storage period; chemicals based on growth regulators promote an increase in emergency velocity rate of the tomato seedlings when applied at pre-sowing at the recommended dose.