Performance and morphology of intestinal mucosa of broilers fed mannan-oligosaccharides and enzymes

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

Production of transgalactosylated oligosaccharides (TOS) by galactosyltransferase activity from Penicillium simplicissimum

Ingestion of transgalactosylated oligosaccharides (TOS) and other non-digestible oligosaccharides (NDOs) induces a significant increase in Bifidobacterium, Lactobacillus and some desirable species of Streptococcus populations in the gut of human and other animals (prebiotic effect). This change in the intestinal flora is responsible for several beneficial physiological effects such as a decrease of putrefactive products in the feces, lower blood cholesterol content, higher Ca2+ absorption, a smaller loss of bone tissue in ovariotomized female rats and a lower incidence of colon cancer. beta-Galactosidase from Penicillium simplicissimum, a strain isolated from soil, showed high galactosyltransferase activity when incubated with a highly concentrated lactose solution. Optimum pH and temperature ranges for hydrolytic activity were 4.0-4.6 and 55-60 degrees C, respectively, for a lactose concentration of 5.0% (w/v). Maximal galactosyltransferase activity was obtained at pH 6.5 and 50 degrees C and TOS synthesis was positively associated with lactose concentration in the reaction medium. Thus, when 50 ml of a 60% (w/v) lactose solution was incubated with 26.6 U of beta-galactosidase under the best pH and temperature conditions for transferase activity, a final product with 30.5% TOS (183 mg ml(-1)), 27.5% residual lactose and 42.0% monosaccharides was obtained. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Production of xylo-oligosaccharides by immobilized-stabilized derivatives of endo-xylanase from Streptomyces halstedii

An endoxylanase from Streptomyces halstedii was stabilized by multipoint covalent immobilization on glyoxyl-agarose supports. The immobilized enzyme derivatives preserved 65% of the catalytic activity corresponding to the one of soluble enzyme that had been immobilized. These immobilized derivatives were 200 times more stable 200 times more stable than the one-point covalently immobilized derivative in experiments involving thermal inactivation at 60 °C. The activity and stability of the immobilized enzyme was higher at pH 5.0 than at pH 7.0. The optimal temperature for xylan hydrolysis was 10 °C higher for the stabilized derivative than for the non-stabilized derivative. On the other hand, the highest loading capacity of activated 10% agarose gels was 75 mg of enzyme per mL of support. To prevent diffusional limitations, low loaded derivatives (containing 0.2 mg of enzyme per mL of support) were used to study the hydrolysis of xylan at high concentration (close to 1% (w/v)). 80% of the reducing sugars were released after 3 h at 55 °C. After 80% of enzymatic hydrolysis, a mixture of small xylo-oligosaccharides was obtained (from xylobiose to xylohexose) with a high percentage of xylobiose and minimal amounts of xylose. The immobilized-stabilized derivatives were used for 10 reaction cycles with no loss of catalytic activity. © 2013 Elsevier Ltd. All rights reserved.

Xylo-oligosaccharides from lignocellulosic materials: Chemical structure, health benefits and production by chemical and enzymatic hydrolysis

Currently, there is worldwide interest in the technological use of agro-industrial residues as a renewable source of food and biofuels. Lignocellulosic materials (LCMs) are a rich source of cellulose and hemicellulose. Hemicellulose is rich in xylan, a polysaccharide used to develop technology for producing alcohol, xylose, xylitol and xylo-oligosaccharides (XOSs). The XOSs are unusual oligosaccharides whose main constituent is xylose linked by β 1-4 bonds. The XOS applications described in this paper highlight that they are considered soluble dietary fibers that have prebiotic activity, favoring the improvement of bowel functions and immune function and having antimicrobial and other health benefits. These effects open a new perspective on potential applications for animal production and human consumption. The raw materials that are rich in hemicellulose include sugar cane bagasse, corncobs, rice husks, olive pits, barley straw, tobacco stalk, cotton stalk, sunflower stalk and wheat straw. The XOS-yielding treatments that have been studied include acid hydrolysis, alkaline hydrolysis, auto-hydrolysis and enzymatic hydrolysis, but the breaking of bonds present in these compounds is relatively difficult and costly, thus limiting the production of XOS. To obviate this limitation, a thorough evaluation of the most convenient methods and the opportunities for innovation in this area is needed. Another challenge is the screening and taxonomy of microorganisms that produce the xylanolytic complex and enzymes and reaction mechanisms involved. Among the standing out microorganisms involved in lignocellulose degradation are Trichoderma harzianum, Cellulosimicrobium cellulans, Penicillium janczewskii, Penicillium echinulatu, Trichoderma reesei and Aspergillus awamori. The enzyme complex predominantly comprises endoxylanase and enzymes that remove hemicellulose side groups such as the acetyl group. The complex has low β-xylosidase activities because β-xylosidase stimulates the production of xylose instead of XOS; xylose, in turn, inhibits the enzymes that produce XOS. The enzymatic conversion of xylan in XOS is the preferred route for the food industries because of problems associated with chemical technologies (e.g., acid hydrolysis) due to the release of toxic and undesired products, such as furfural. The improvement of the bioprocess for XOS production and its benefits for several applications are discussed in this study. © 2012 Elsevier Ltd.

Protective effect of mannan oligosaccharides against early colonization by Salmonella Enteritidis in chicks is improved by higher dietary threonine levels

Aims: To evaluate mannan oligosaccharide (MOS) and threonine effects on performance, small intestine morphology and Salmonella spp. counts in Salmonella Enteritidis-challenged birds. Methods and Results: One-day-old chicks (1d) were distributed into five treatments: nonchallenged animals fed basal diet (RB-0), animals fed basal diet and infected with Salmonella Enteritidis (RB-I), animals fed high level of threonine and infected (HT-I), birds fed basal diet with MOS and infected (MOS-I), birds fed high level of threonine and MOS and infected (HT+MOS-I). Birds were inoculated at 2d with Salmonella Enteritidis, except RB-0 birds. Chicks fed higher dietary threonine and MOS showed performance similar to RB-0 and intestinal morphology recovery at 8 dpi. Salmonella counts and the number of Salmonella-positive animals were lower in HT+MOS-I compared with other challenged groups. Conclusion: Mannan oligosaccharides and threonine act synergistically, resulting in improved intestinal environment and recovery after Salmonella inoculation. Significance and Impact of the Study: Nutritional approaches may be useful to prevent Salmonella infection in the first week and putative carcass contamination at slaughter. This is the first report on the possible synergistic effect of mannan oligosaccharides and threonine, and further studies should be performed including performance, microbiota evaluation, composition of intestinal mucins and immune assessment. © 2012 The Society for Applied Microbiology.

Enzymatic production of functional oligosaccharides - A review

Interest in oligosaccharide production and its general characteristics is growing. The physiological effects resulting from its ingestion make them more attractive than its sweetness, and the versatility of these carbohydrates allows their use for human and animal nutrition, pharmacology and the cosmetics industry, among others. Several microorganisms are involved in enzyme production to create oligomers with biological activity, including fructooligosaccharides, galactooligosaccharides and aminoglucanoligosaccharides. Some oligomers are currently on the market, but the search for new microorganisms producing enzymes, high-yield processes for obtaining oligosaccharides, different physiological effects, and a correlation between chemical structure and function continues.

Production of β-(1,3)-glucanases by Trichoderma harzianum Rifai: Optimization and Application to Produce Gluco-oligosaccharides from Paramylon and Pustulan

β-(1→3)-Glucanases were produced by Trichoderma harzianum Rifai PAMB-86 cultivated on botryosphaeran in a bench-fermenter and optimised by the response surface method. Maximal enzyme titres occurred at 5 days, initial pH 5.5 and aeration of 1.5vvm. β-(1→3)-The β-glucanolytic enzyme complex produced by T. harzianum Rifai PAMB- 86 was fractionated by gel filtration into 2 fractions (F-I, F-II), and employed to produce gluco-oligosaccharides from algal paramylon ((1→3)-β-D-glucan) and lichen pustulan ((1→6)-β-D-glucan). Both enzymes attacked paramylon to the extent of ~15-20% in 30 min releasing glucose and laminaribiose as major end-products, and laminarioligosaccharides of degree of polymerization (DP) ≥3. Only F-I degraded pustulan resulting in ~2% degradation at 30 min, with glucose, gentiobiose and gentio-oligosaccharides of DP ≥4 as major products. The difference in the nature of the hydrolysis products can be explained by the substrate specificities of each enzyme fraction, and the structural differences of the β-D-glucans attacked.

Utilização de nutrientes de dietas contendo mananoligossacarídeo e/ou complexo enzimático para frangos de corte

Avaliaram-se a digestibilidade ileal e a retenção de alguns nutrientes e os valores de energia de dietas contendo mananoligossacarídeo (MOS) e/ou complexo enzimático (CE) para frangos de corte. Foram utilizadas 275 aves em delineamento em blocos ao acaso e arranjo fatorial 2 x 2 + 1, com dois níveis de MOS (0 e 0,1%), dois níveis de complexo enzimático (0 e 0,05%) e uma dieta controle positivo com antibióticos. O óxido crômico (0,5%) foi adicionado às dietas para estimativa do fator de indigestibilidade. O experimento teve início quando as aves completaram 13 dias de idade; a coleta de excretas foi realizada do 20º ao 22º dia e a de digesta, no 23º dia de idade das aves. A interação MOS × CE foi significativa para a retenção de PB e P e de energia metabolizável aparente (EMA), cujos valores foram maiores nas dietas com MOS e CE. A inclusão do CE melhorou a retenção de MS e os coeficientes de digestibilidade ileal de MS, PB, Ca e P na retenção de cálcio e nos valores de energia digestível com a inclusão de mananoligossacarídeo. Os coeficientes de digestibilidade ileal da MS, a retenção de MS, PB, Ca e P e os valores de energia digestível e de EMA das dietas contendo MOS e/ou CE foram superiores aos obtidos com a dieta contendo antibióticos.

Mananoligossacarídeos e complexo enzimático em dietas para frangos de corte

Avaliou-se o efeito de dietas com mananoligossacarídeos e complexo enzimático (CE) sobre o desempenho, a morfologia intestinal e a qualidade da cama de frangos aos 42 dias de idade. Foram utilizadas 750 aves em delineamento inteiramente casualizado em esquema fatorial 2 × 2 + 1, com dois níveis de mananoligossacarídeos (0 e 0,1% de 1 a 21 dias e 0,05% de 22 a 42 dias de idade), dois níveis de complexo enzimático (0 e 0,05%) e uma dieta com antibióticos (CP), totalizando cinco dietas com cinco repetições. Aos 42 dias de criação, o desempenho foi avaliado e, após o abate das aves, foram coletadas amostras de intestino e de cama e avaliado o desempenho. A inclusão de mananoligossacarídeos e/ou complexo enzimático na dieta não afetou o desempenho das aves, o perímetro e a altura dos vilos duodenais, a profundidade de criptas, a densidade de vilos no duodeno, jejuno e íleo, os teores de matéria seca e nitrogênio total e o pH das camas. A interação mananoligossacarídeos × complexo enzimático foi significativa para perímetro e altura de vilos no jejuno, que foram maiores nas aves alimentadas com as rações sem complexo enzimático e mananoligossacarídeos, mesmo comportamento observado para perímetro e altura de vilos ileais. Entretanto, quando adicionados mananoligossacarídeos e complexo enzimático, os valores dessas variáveis reduziram. A volatilização de amônia aumentou em camas de frango tratados com antibióticos e diminuiu com a adição de mananoligossacarídeos à dieta. A adição de mananoligossacarídeos ou complexo enzimático às dietas aumentou o perímetro e altura de vilos da mucosa do jejuno e do íleo e reduziu a volatilização de amônia da cama.

Mananoligossacarídeos em dietas para frangos de corte

Objetivou-se avaliar dietas contendo mananoligossacarídeos (MOS) como aditivo alternativo aos promotores de crescimento por meio do estudo da morfometria do intestino e do desempenho de frangos de corte. Para tanto, 1280 pintos de corte foram distribuídos em delineamento inteiramente casualizado com quatro tratamentos (controle negativo, CN: dieta isenta de antibiótico; controle positivo, CP: dieta contendo antibiótico e duas dietas, MOS 1 e MOS 2, nas quais foram adicionadas ao CN duas fontes distintas de MOS) e oito repetições, sendo a unidade experimental composta por 40 aves. Para submeter as aves ao desafio sanitário, foi formulada uma dieta basal com milho, farelo de soja e farinha de carne e ossos. Adotou-se cama reutilizada, limpeza dos bebedouros duas vezes por semana e oferta semanal de água contaminada com cama. Foram avaliadas altura de vilo e profundidade de cripta do duodeno, jejuno e íleo, consumo da dieta, peso médio, ganho de peso e conversão alimentar das aves. Houve melhora na profundidade de cripta no jejuno e na altura de vilo no íleo das aves alimentadas com dietas contendo MOS. A adição de MOS, independente da fonte, resultou em melhor conversão alimentar em relação às aves do CN, sendo similares às aves do CP. Os mananoligossacarídeos podem ser utilizados como aditivo alternativo aos promotores de crescimento em dietas para frangos de corte, porém, dependendo da fonte, esta pode acarretar em pequenas diferenças no desempenho das aves.

Effect of probiotic and prebiotic inclusion in weaned piglet diets on structure and ultra-structure of small intestine

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

Properties of a new fungal b-galactosidase with potential application in the dairy industry

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

Enzymatic hydrolysis of botryosphaeran and laminarin by beta-1,3-glucanases produced by Botryosphaeria rhodina and Trichoderma harzianum Rifai

Botryosphaeran, a (1 -> 3; 1 -> 6)-beta-D-glucan produced by Botryosphaeria rhodina, and laminarin were hydrolysed by two fungal beta-glucanases predominantly of the 1,3-type produced by B. rhodina and Trichoderma harzianum Rifai grown on botryosphaeran as sole carbon source. Both beta-glucanase preparations presented different modes of attack on botryosphaeran and laminarin. Laminarin was hydrolysed to the extent of similar to 50% in 1 hand 100% within 24 h, and its hydrolysis products were mainly glucose and gentiobiose, and lesser amounts of laminaribiose and oligosaccharides of DP 3-4 during the early stages of hydrolysis, while botryosphaeran 'yielded mainly glucose and gentiobiose with some trisaccharide, but no laminaribiose or tetrasaccharide when hydrolysed by the T. harzianum enzyme. By contrast, B. rhodina beta-1,3-glucanases produced predominantly glucose during all stages of botryosphaeran hydrolysis. Some physicochemical properties of the 1,3- and 1,6-beta-glucanases, and beta-glucosidases contained in the two fungal P-glucanase preparations are also described for the first time. (c) 2006 Elsevier Ltd. All rights reserved.

Obtenção de oligossacarídeos N-ligados às glicoproteínas dos líquens Sticta tomentosa e Sticta damaecornis

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