Carbohydrases and Phytase in Poultry and Pig Nutrition: A Review beyond the Nutrients and Energy Matrix.

This review aimed to clarify the mechanisms through which exogenous enzymes (carbohydrases and phytase) influence intestinal health, as well as their effects on the nutrients and energy matrix in diets fed to poultry and pigs reared under sanitary challenging conditions. Enzyme supplementation can positively affect intestinal microbiota, immune system, and enhance antioxidant status. Although enzymes have been shown to save energy and nutrients, their responses under sanitary challenging conditions are poorly documented. Immune system activation alters nutrient partitioning, which can affect the matrix values for exogenous enzymes on commercial farms. Notably, the carbohydrases and phytase supplementation under sanitary challenging conditions align with energy and nutritional valorization matrices. Studies conducted under commercial conditions have shown that matrices containing carbohydrases and phytase can maintain growth performance and health in poultry and pigs. However, these studies have predominantly focused on assessing a single level of reduction in energy and/or available phosphorus and total calcium, limiting our ability to quantify potential energy and nutrient savings in the diet. Future research should delve deeper into determining the extent of energy and nutrient savings and understanding the effects of alone or blended enzymes supplementation to achieve more specific insights.

Complexo enzimático em rações para leitões nas fases pré-inicial e inicial / Enzymatic complex in piglet feeds in the pre-initial and initial phases

Objetivou-se, com este trabalho, avaliar o efeito da utilização de complexo enzimático em rações para leitões sobre o desempenho zootécnico nas fases pré-inicial e inicial. Foram utilizados, para isso, 292 leitões, machos castrados e fêmeas, de linhagem comercial. O período de suplementação da enzima, na maternidade, foi do sétimo ao vigésimo primeiro dia de idade, e, na creche, foi do desmame até a saída para a fase de crescimento. O delineamento utilizado foi inteiramente casualizado, sendo a matriz e sua leitegada consideradas a unidade experimental durante a maternidade e uma baia com dez leitões na creche. Na maternidade, os tratamentos consistiram em: T1 - ração controle, sem complexo enzimático; T2 - ração controle com adição on top do complexo enzimático com inclusão de 5%. Na creche, foram definidos três tratamentos: T1 - ração controle; T2 - ração controle com inclusão on top de 5% do complexo enzimático na fase de maternidade; T3 - ração controle com inclusão de 5% de complexo enzimático nas fases de maternidade e creche. A suplementação durante a lactação não influenciou o desempenho da leitegada na maternidade até o 21º dia. Na creche, leitões alimentados com ração controle apresentaram maior peso inicial em relação aos suplementados na fase de maternidade e creche. Ao término da fase, os leitões mostraram desempenho semelhante, independente da dieta. Conclui-se que a inclusão do complexo enzimático na forma on top em dietas para leitões não influenciou parâmetros de desempenho na maternidade, porém, favoreceu a recuperação de leitões desmamados com menor peso.

This study aimed to evaluate the effect of using an enzymatic complex in piglet feeds on the zootechnical performance in the pre-starter and initial phases. For this purpose, 292 piglets, castrated males and females, of commercial lineage were used. The enzyme supplementation period in the maternity hospital was from the seventh to the twenty-first day of age, and in the nursery, it was from the waning until the exit for the growth phase. The design used was completely randomized, with the mother and her litter considered the experimental unit during maternity and a stall with 10 piglets in the nursery. In maternity, the treatments consisted of: T1 - control feed, without the enzymatic complex; T2 - control diet with on top addition of the enzymatic complex with 5% inclusion. Three treatments were defined at the nursery: T1 - control feed; T2 - control diet with inclusion on top of 5% of the enzymatic complex in the maternity phase; T3 - control feed with the inclusion of 5% enzymatic complex in the maternity and nursery phases. Supplementation during lactation did not influence litter performance in the maternity unit until 21 days. In the nursery, piglets fed with control feed had higher initial weight compared to those supplemented in the maternity and nursery phases. At the end of the phase, the piglets showed similar performance, regardless of diet. It is concluded that the inclusion of the enzymatic complex used in the on top form in diets for piglets did not influence performance parameters in maternity; however, it favored the recovery of weaned piglets with lower weight.

Enzimas exógenas na dieta de leitões desmamados / Exogenous enzymes in the diet of weaned piglets

O desmame é caracterizado como o período mais crítico da produção de suínos devido à imaturidade digestiva e à baixa concentração de enzimas digestivas, como carboidrases, proteases e fitases, nas primeiras semanas do pós-desmame, o que ocasiona o baixo desempenho dos leitões. Neste caso, há a necessidade de suplementação exógena de enzimas que atuam na degradação de frações do alimento, aumentando o aproveitamento da dieta. A nutrição enzimática exógena tem sido considerada uma alternativa eficaz frente à baixa concentração de enzimas endógenas nas primeiras semanas após o desmame por contribuir para o aumento na digestibilidade das dietas sólidas e, deste modo, para um maior aproveitamento dos ingredientes. Na nutrição de suínos, as enzimas exógenas mais utilizadas são as carboidrases, as proteases e as fitases. Ademais, a utilização de blends enzimáticos também é considerada uma ótima alternativa, pois contribui para a inibição de fatores antinutricionais, os quais são responsáveis por indisponibilizar nutrientes necessários para o desenvolvimento e desempenho dos animais. Diante do exposto, o objetivo deste estudo foi apresentar as principais enzimas exógenas utilizadas na nutrição de leitões desmamados, bem como seus efeitos no desempenho animal.

Weaning is characterized as the most critical period in swine production due to digestive immaturity and low concentration of digestive enzymes, such as carbohydrases, proteases, and phytases, in the first weeks after weaning, which causes poor performance of piglets. In this case, there is the need for exogenous supplementation of enzymes that act on the degradation of food fractions, increasing diet utilization. Exogenous enzyme nutrition has been considered an effective alternative to the low concentration of endogenous enzymes in the first weeks after weaning, as it contributes to an increase in the digestibility of solid diets and, consequently, greater use of the ingredients. In swine nutrition, the most used exogenous enzymes are carbohydrases, proteases, and phytases. Furthermore, the use of enzymatic blends is also considered a great alternative, as they contribute to the inhibition of anti-nutritional factors, which are responsible for making nutrients necessary for the development and performance of animals unavailable. Therefore, this study aimed to present the main exogenous enzymes used in the nutrition of weaned piglets, as well as their effects on animal performance.

A proteomic approach to identify digestive enzymes, their exocytic and microapocrine secretory routes and their compartmentalization in the midgut of Spodoptera frugiperda.

A proteomic approach was used to identify the digestive enzymes secreted by exocytosis and by microapocrine vesicles and enzyme midgut compartmentalization in Spodoptera frugiperda larvae. For this, proteomic analyses were performed in isolated midgut enterocyte microvillar membrane, in a fraction enriched in microapocrine vesicles (separated in soluble and membrane fractions), in the washings of the peritrophic membrane to isolate its loosely- and tightly-bound proteins, and in the peritrophic membrane contents. PM washings correspond to proteins extracted from the mucus layer surrounding PM. Serine endopeptidases (trypsins, chymotrypsins and serine endopeptidase homologs that have substitutions in the catalytic residues) and lipases are mainly secreted by exocytosis. Aminopeptidases are mainly microvillar enzymes and some are secreted membrane-bound to microapocrine vesicles, whereas carboxypeptidase isoforms follow different secretory routes. The results also showed that most polymer hydrolases (such as amylase and endopeptidases) are not retained in the ectoperitrophic fluid (found in PM washings but absent from PM contents). On the contrary, most enzymes involved in intermediate digestion (exemplified by carboxypeptidase and aminopeptidase) do not pass through the peritrophic membrane. Finally, the data revealed that the protein composition of PM includes peritrophins classified as peritrophic membrane proteins, PMP, and chitin deacetylase.

A proteomic approach to identify digestive enzymes, their exocytic and microapocrine secretory routes and their compartmentalization in the midgut of Spodoptera frugiperda

A proteomic approach was used to identify the digestive enzymes secreted by exocytosis and by microapocrine vesicles and enzyme midgut compartmentalization in Spodoptera frugiperda larvae. For this, proteomic analyses were performed in isolated midgut enterocyte microvillar membrane, in a fraction enriched in microapocrine vesicles (separated in soluble and membrane fractions), in the washings of the peritrophic membrane to isolate its loosely- and tightly-bound proteins, and in the peritrophic membrane contents. PM washings correspond to proteins extracted from the mucus layer surrounding PM. Serine endopeptidases (trypsins, chymotrypsins and serine endopeptidase homologs that have substitutions in the catalytic residues) and lipases are mainly secreted by exocytosis. Aminopeptidases are mainly microvillar enzymes and some are secreted membrane-bound to microapocrine vesicles, whereas carboxypeptidase isoforms follow different secretory routes. The results also showed that most polymer hydrolases (such as amylase and endopeptidases) are not retained in the ectoperitrophic fluid (found in PM washings but absent from PM contents). On the contrary, most enzymes involved in intermediate digestion (exemplified by carboxypeptidase and aminopeptidase) do not pass through the peritrophic membrane. Finally, the data revealed that the protein composition of PM includes peritrophins classified as peritrophic membrane proteins, PMP, and chitin deacetylase.

Addition of a blend of exogenous enzymes to broiler chickens diets: impacts on performance and production costs / Adição de uma mistura de enzimas exógenas a dietas de frangos de corte: impactos no desempenho e custos de produção

Broiler diets are based on corn, soybean and wheat production; however, some protein ingredients have many antinutritional factors and low digestibility. The objective of this study was to add a blend of exogenous enzymes to the feed with low nutritional value for broilers to reduce production costs and improve digestibility while maintaining good zootechnical performance. The experimental design was completely randomized, including three treatments with five replications (n=15) each: a) positive control (PC), diet calculated for males with average performance; b) negative control + exogenous enzymes (NC+EE), minimum nutrient diet according to the requirements of the production phase, and the enzymatic blend was added; and c) negative control (NC), minimum levels of nutrients for each phase. At 21 days, the PC group showed greater weight gain and lower feed conversion than the NC (P<0.05). At 42 days, PC had lower feed intake than NC (P=0.040), while lower feed conversion was observed in groups PC and NC+EE than NC (P=0.001). The production efficiency index was higher in the PC treatment, but the NC+EE treatment was higher than the NC (P=0.001). Considering production costs and body weight, we found that NC+EE birds had greater profitability. Therefore, we conclude that the blend of exogenous enzymes added to a diet with minimal nutritional levels has practical application in the broiler production system.(AU)

As dietas de frangos de corte são baseadas na produção de milho, soja e trigo; no entanto, alguns ingredientes proteicos possuem muitos fatores antinutricionais e baixa digestibilidade. O objetivo deste estudo foi adicionar à ração uma mistura de enzimas exógenas com baixo valor nutricional para frangos de corte para reduzir os custos de produção e melhorar a digestibilidade, mantendo o bom desempenho zootécnico. O delineamento experimental foi inteiramente casualizado, incluindo três tratamentos com cinco repetições (n=15) cada: a) controle positivo (PC), dieta calculada para machos com desempenho médio; b) controle negativo + enzimas exógenas (CN+EE), dieta mínima de nutrientes de acordo com as exigências da fase de produção, e foi adicionada a mistura enzimática; c) controle negativo (CN), teores mínimos de nutrientes para cada fase. Aos 21 dias, o grupo PC apresentou maior ganho de peso e menor conversão alimentar que o NC (P<0,05). Aos 42 dias, PC apresentou menor consumo de ração que NC (P=0,040), enquanto foi observada menor conversão alimentar nos grupos PC e NC+EE que NC (P=0,001). O índice de eficiência de produção foi maior no tratamento PC, mas o tratamento NC+EE foi maior que o NC (P=0,001). Considerando os custos de produção e peso corporal, verificamos que as aves NC+EE tiveram maior rentabilidade. Portanto, concluímos que a mistura de enzimas exógenas adicionada a uma dieta com níveis nutricionais mínimos tem aplicação prática no sistema de produção de frangos de corte.(AU)

Use of different combinations of enzyme complexes in broiler diets / Utilização de diferentes combinações de complexos enzimáticos em rações para frangos de corte

Two experiments were carried out to investigate the effect of "on top" addition of different enzyme complexes, the enzyme α-galactosidase and three sources of the enzyme phytase available on the market, in broiler diets. In the first experiment, 1260 one-day-old Cobb 500® chicks were distributed into seven treatments in a completely randomized design (CRD) with six replicates and 30 birds/replicate. Treatments consisted of combinations of different enzyme complexes, namely, complex A (phytase, protease, xylanase, ß-glucanase, cellulase, amylase, pectinase), complex B (protease and cellulase) and complex C (xylanase, amylase and protease); isolated α-galactosidase (GAL); and three sources of phytase (P1, P2 and P3) in the diet. The treatments were formulated as follows: T1 - basal diet (BD); T2 - BD + enzyme complex A + enzyme complex B (BDAB); T3 - BDAB + GAL; T4 - BD + complex A + GAL; T5 - BD + complex C + P1 + GAL (BDCG); T6- BDCG + P2; and T7 - BDCG + P3. The following variables were measured in the experimental period of 42 days: feed intake (FI), weight gain (WG), average final weight (AFW), feed conversion (FC), and carcass yield. Significant differences occurred for AFW, WG and FC in the pre-starter phase. In the second experiment,112 Cobb 500® chicks aged 25 days were distributed into seven treatments in a CRD with four replicates and four birds/replicate. Treatments were the same as in the first experiment. Nutrient digestibility was evaluated in an experimental period of seven days. Differences were found in the metabolism coefficient of ether extract (MCEE). Dietary inclusion of enzyme complexes improves the AFW and WG of chickens from1 to 7 days of age and MCEE in the grower phase.(AU)

Foram realizados dois experimentos com o objetivo de avaliar o efeito da adição "on top" de diferentes complexos enzimáticos, enzima α-galactosidade e três fontes de enzima fitase disponíveis no mercado em dietas para frangos de corte. No primeiro experimento 1260 pintos Cobb 500® com um dia de idade foram distribuídos em delineamento inteiramente casualizado (DIC), sete tratamentos, seis repetições e 30 aves/repetição. Tratamentos consistiram na associação de diferentes complexos enzimáticos: complexo A (fitase, protease, xilanase, ß-glucanase, celulase, amilase, pectinase); complexo B (protease e celulase); e complexo C (xilanase, amilase e protease); α-galactosidase isolada (GAL); e três fontes de fitase (F1, F2 e F3) nas rações. Em que: T1 - ração basal (RB); T2 - RB + complexo enzimático A + complexo enzimático B (RBAB); T3 - RBAB + GAL; T4 - RB + complexo A + GAL; T5 - RB + complexo C + F1 + GAL (RBCG); T6 - RBCG+F2; T7 - RBCG + F3. O período experimental foi de 42 dias. Avaliaram-se: consumo de ração (CR), ganho de peso (GP), peso médio final (PMF), conversão alimentar (CA), e rendimento de carcaça. Foram observadas diferenças significativas para PMF, GP e CA na fase pré-inicial. No segundo experimento 112 pintos Cobb 500® com 25 dias de idade foram distribuídos em DIC, sete tratamentos, quatro repetições e quatro aves/repetição. Os tratamentos foram os mesmos do primeiro experimento. O período experimental foi de sete dias, avaliou-se a digestibilidade dos nutrientes. Foram observadas diferenças para coeficiente de metabolizabilidade do extrato etéreo (CMEE). A inclusão de complexos enzimáticos nas rações melhora o PMF e GP de frangos de 1 a 7 dias de idade e o CMEE na fase de crescimento.(AU)

Xylanases in diets with alternative feeds and energy reduction in commercial layers / Xilanases em dietas com alimentos alternativos e redução energética para poedeiras comerciais

The use of alternative ingredients has been increasing in a continuing attempt to reduce production costs, along with the use of additives, such as carbohydrases, for their possible positive effects on nutrient metabolization by layers. Thus, this study aimed to evaluate the effects of different xylanases in diets with reduced metabolizable energy (ME) and the inclusion of alternative ingredients on the metabolizability of nutrients in diets for commercial laying hens, by conducting two metabolism tests. In the first trial, 100 layers hens were distributed in a fully randomized 2 × 2 + 1 factorial design (two metabolizable energy reductions × two xylanases, plus one control diet). In the second assay, 140 layer hens were distributed in a fully randomized 2 × 3 + 2 factor design (two xylanases× three ingredients, plus two control diets, positive and negative). It was concluded that supplementation with pantanal xylanase improved the metabolizability of nutrients in corn-based diets and soybean meal for light commercial layers, allowing for a reduction of up to 200 kcal/kg of ME in the diets. Xylanases are more effective on wheat bran enabling a decrease of 150 kcal/kg of ME in commercial laying diets without affecting nutrient metabolizability. The action of pantanal xylanase in diets containing fibrous ingredients was similar to that of commercial xylanase.(AU)

O uso de ingredientes alternativos vem aumentando em uma tentativa contínua de reduzir os custos de produção, juntamente com o uso de aditivos, como os carboidrases, para seus possíveis efeitos positivos na metabolização de nutrientes por poedeiras. Assim, este estudo teve como objetivo avaliar os efeitos de diferentes xilanases em dietas com energia metabolizável reduzida (ME) e a inclusão de ingredientes alternativos sobre a metabolizabilidade de nutrientes em dietas para galinhas poedeiras comerciais, realizando dois testes de metabolismo. No primeiro ensaio, 100 aves foram distribuídas em delineamento inteiramente casualizado em esquema fatorial 2 x 2 + 1 (duas reduções de energia metabolizável x duas xilanases, mais uma dieta controle). No segundo ensaio, 140 aves foram distribuídas em delineamento inteiramente casualizado em esquema fatorial 2 x 3 + 2 (duas xilanases x três ingredientes -, mais duas dietas controle positivo e controle negativo). Conclui-se que a suplementação com xilanase do Pantanal melhora a metabolizabilidade dos nutrientes de dietas a base de milho e farelo de soja para poedeiras comerciais leves, possibilitando a redução em até 200 Kcal/kg de energia metabolizável das dietas. As xilanases são mais efetivas sobre o farelo de trigo e possibilitam a redução de 150 Kcal/kg de energia metabolizável nas dietas de poedeiras comerciais, sem prejuízo na metabolizabilidade dos nutrientes. Em dietas contendo ingredientes fibrosos a ação da xilanase do Pantanal é similar a xilanase comercial.(AU)

Use of different combinations of enzyme complexes in broiler diets / Utilização de diferentes combinações de complexos enzimáticos em rações para frangos de corte

Two experiments were carried out to investigate the effect of "on top" addition of different enzyme complexes, the enzyme α-galactosidase and three sources of the enzyme phytase available on the market, in broiler diets. In the first experiment, 1260 one-day-old Cobb 500® chicks were distributed into seven treatments in a completely randomized design (CRD) with six replicates and 30 birds/replicate. Treatments consisted of combinations of different enzyme complexes, namely, complex A (phytase, protease, xylanase, ß-glucanase, cellulase, amylase, pectinase), complex B (protease and cellulase) and complex C (xylanase, amylase and protease); isolated α-galactosidase (GAL); and three sources of phytase (P1, P2 and P3) in the diet. The treatments were formulated as follows: T1 - basal diet (BD); T2 - BD + enzyme complex A + enzyme complex B (BDAB); T3 - BDAB + GAL; T4 - BD + complex A + GAL; T5 - BD + complex C + P1 + GAL (BDCG); T6- BDCG + P2; and T7 - BDCG + P3. The following variables were measured in the experimental period of 42 days: feed intake (FI), weight gain (WG), average final weight (AFW), feed conversion (FC), and carcass yield. Significant differences occurred for AFW, WG and FC in the pre-starter phase. In the second experiment,112 Cobb 500® chicks aged 25 days were distributed into seven treatments in a CRD with four replicates and four birds/replicate. Treatments were the same as in the first experiment. Nutrient digestibility was evaluated in an experimental period of seven days. Differences were found in the metabolism coefficient of ether extract (MCEE). Dietary inclusion of enzyme complexes improves the AFW and WG of chickens from1 to 7 days of age and MCEE in the grower phase.

Foram realizados dois experimentos com o objetivo de avaliar o efeito da adição "on top" de diferentes complexos enzimáticos, enzima α-galactosidade e três fontes de enzima fitase disponíveis no mercado em dietas para frangos de corte. No primeiro experimento 1260 pintos Cobb 500® com um dia de idade foram distribuídos em delineamento inteiramente casualizado (DIC), sete tratamentos, seis repetições e 30 aves/repetição. Tratamentos consistiram na associação de diferentes complexos enzimáticos: complexo A (fitase, protease, xilanase, ß-glucanase, celulase, amilase, pectinase); complexo B (protease e celulase); e complexo C (xilanase, amilase e protease); α-galactosidase isolada (GAL); e três fontes de fitase (F1, F2 e F3) nas rações. Em que: T1 - ração basal (RB); T2 - RB + complexo enzimático A + complexo enzimático B (RBAB); T3 - RBAB + GAL; T4 - RB + complexo A + GAL; T5 - RB + complexo C + F1 + GAL (RBCG); T6 - RBCG+F2; T7 - RBCG + F3. O período experimental foi de 42 dias. Avaliaram-se: consumo de ração (CR), ganho de peso (GP), peso médio final (PMF), conversão alimentar (CA), e rendimento de carcaça. Foram observadas diferenças significativas para PMF, GP e CA na fase pré-inicial. No segundo experimento 112 pintos Cobb 500® com 25 dias de idade foram distribuídos em DIC, sete tratamentos, quatro repetições e quatro aves/repetição. Os tratamentos foram os mesmos do primeiro experimento. O período experimental foi de sete dias, avaliou-se a digestibilidade dos nutrientes. Foram observadas diferenças para coeficiente de metabolizabilidade do extrato etéreo (CMEE). A inclusão de complexos enzimáticos nas rações melhora o PMF e GP de frangos de 1 a 7 dias de idade e o CMEE na fase de crescimento.

Xylanases in diets with alternative feeds and energy reduction in commercial layers / Xilanases em dietas com alimentos alternativos e redução energética para poedeiras comerciais

The use of alternative ingredients has been increasing in a continuing attempt to reduce production costs, along with the use of additives, such as carbohydrases, for their possible positive effects on nutrient metabolization by layers. Thus, this study aimed to evaluate the effects of different xylanases in diets with reduced metabolizable energy (ME) and the inclusion of alternative ingredients on the metabolizability of nutrients in diets for commercial laying hens, by conducting two metabolism tests. In the first trial, 100 layers hens were distributed in a fully randomized 2 × 2 + 1 factorial design (two metabolizable energy reductions × two xylanases, plus one control diet). In the second assay, 140 layer hens were distributed in a fully randomized 2 × 3 + 2 factor design (two xylanases× three ingredients, plus two control diets, positive and negative). It was concluded that supplementation with pantanal xylanase improved the metabolizability of nutrients in corn-based diets and soybean meal for light commercial layers, allowing for a reduction of up to 200 kcal/kg of ME in the diets. Xylanases are more effective on wheat bran enabling a decrease of 150 kcal/kg of ME in commercial laying diets without affecting nutrient metabolizability. The action of pantanal xylanase in diets containing fibrous ingredients was similar to that of commercial xylanase.

O uso de ingredientes alternativos vem aumentando em uma tentativa contínua de reduzir os custos de produção, juntamente com o uso de aditivos, como os carboidrases, para seus possíveis efeitos positivos na metabolização de nutrientes por poedeiras. Assim, este estudo teve como objetivo avaliar os efeitos de diferentes xilanases em dietas com energia metabolizável reduzida (ME) e a inclusão de ingredientes alternativos sobre a metabolizabilidade de nutrientes em dietas para galinhas poedeiras comerciais, realizando dois testes de metabolismo. No primeiro ensaio, 100 aves foram distribuídas em delineamento inteiramente casualizado em esquema fatorial 2 x 2 + 1 (duas reduções de energia metabolizável x duas xilanases, mais uma dieta controle). No segundo ensaio, 140 aves foram distribuídas em delineamento inteiramente casualizado em esquema fatorial 2 x 3 + 2 (duas xilanases x três ingredientes -, mais duas dietas controle positivo e controle negativo). Conclui-se que a suplementação com xilanase do Pantanal melhora a metabolizabilidade dos nutrientes de dietas a base de milho e farelo de soja para poedeiras comerciais leves, possibilitando a redução em até 200 Kcal/kg de energia metabolizável das dietas. As xilanases são mais efetivas sobre o farelo de trigo e possibilitam a redução de 150 Kcal/kg de energia metabolizável nas dietas de poedeiras comerciais, sem prejuízo na metabolizabilidade dos nutrientes. Em dietas contendo ingredientes fibrosos a ação da xilanase do Pantanal é similar a xilanase comercial.

Evolution of World and Brazilian Markets for Enzymes Produced by Solid-state Fermentation: A Patent Analysis.

BACKGROUND: The use of enzymes in various industrial processes has become increasingly frequent. When added to productive processes, it can accelerate reactions and generate a number of new products. The solid state fermentation (SSF), among other applications, has been employed also to obtain enzymes. OBJECTIVE: The purpose of this prospection was to map registered patent documents about enzymes production by this type of fermentation in the world, identify the most obtained enzymes with patent documents and compilate information about the world and Brazilian enzyme markets. METHODS: The experimental design was carried out by the keyword-driven scope through the advanced search in the Espacenet database European Patent Office (EPO). The keywords selected were solid-state fermentation and the International Patent Classification code, C12N9 (enzymes; proenzymes), for prospecting of interest. RESULTS: In 2012, there was the higher number of registered patents (12). China holds 84% of deposited patents. Among the types of depositors, 54% of the selected patent documents were deposited by universities and institutes, and 44% by companies. 76.5% of the evaluated patents used fungi as enzyme producer. Analyzing the enzymes obtained in the registered patents, it is verified that the majority belongs to the group of carbohydrases with 43%, followed by proteases (25%), which are also the two classes of enzymes most commercialized in the market. CONCLUSION: China holds the majority of the registered patents but North America gets the largest global enzyme market revenue followed by Europe and Pacific Asia. Carbohydrases were the most commercialized enzymes and with the highest number of patents registered. Among the carbohydrases, cellulases, xylanases and amylases are the most frequent in patent registration while being fungi produced.

Effects of the Dietary Inclusion of Xylanase on the Performance and Jejunum Morphometry of Meat-Type Quails

The objective of this study was to evaluate the effects of the supplementation of two xylanase products to diets with reduced metabolizable energy fed to meat-type quails during the starter phase (1-14 days). A completely randomized experimental design in a 2 x 3 + 1 factorial arrangement two reduced metabolizable energy (ME) diets, inclusion or not of xylanase, and a control diet with no enzyme addition) was applied, totaling seven treatments with five replicates of 45 quails each. At 14 days of age, jejunum segments were collected for morphometry evaluation. No interaction between the studied factors were detected for performance and jejunal morphometry parameters. Body weight, weight gain, feed intake and feed conversion were not influenced by enzyme inclusion. Reduced ME diets (-70 or -140 kcal/kg) did not affect performance, except for feed intake. Xylanase inclusion increased villus height and villus:crypt ratio. Therefore, xylanase supplementation can be effective in corn and soybean meal-based diets, without causing any impairment in the performance of 1- to 14-day-old quails. Xylanases A and B were more efficient when dietary energy level was reduced in 140 kcal ME/kg, and were also shown to effectively improve the jejunal morphometry of starter meat-type quails.(AU)

Effects of the Dietary Inclusion of Xylanase on the Performance and Jejunum Morphometry of Meat-Type Quails

The objective of this study was to evaluate the effects of the supplementation of two xylanase products to diets with reduced metabolizable energy fed to meat-type quails during the starter phase (1-14 days). A completely randomized experimental design in a 2 x 3 + 1 factorial arrangement two reduced metabolizable energy (ME) diets, inclusion or not of xylanase, and a control diet with no enzyme addition) was applied, totaling seven treatments with five replicates of 45 quails each. At 14 days of age, jejunum segments were collected for morphometry evaluation. No interaction between the studied factors were detected for performance and jejunal morphometry parameters. Body weight, weight gain, feed intake and feed conversion were not influenced by enzyme inclusion. Reduced ME diets (-70 or -140 kcal/kg) did not affect performance, except for feed intake. Xylanase inclusion increased villus height and villus:crypt ratio. Therefore, xylanase supplementation can be effective in corn and soybean meal-based diets, without causing any impairment in the performance of 1- to 14-day-old quails. Xylanases A and B were more efficient when dietary energy level was reduced in 140 kcal ME/kg, and were also shown to effectively improve the jejunal morphometry of starter meat-type quails.

Xylanase for meat-type quails from 15 to 35 days old

The objective of this study was to evaluate the effect of the supplementation of two xylanases in the diet of meat-type quail on performance, morphology of the intestinal mucosa, metabolization of nutrients, and carcass yield. The experimental design was a completely randomized 2×3+1 factorial arrangement [two metabolisable energy reductions (70 and 140 kcal/kg), with or without the inclusion of two xylanases (A and B), plus a control treatment without xylanase), totalling seven treatments with five replicates and 42 quail each. Total excreta were collected at 28 days of age to determine the metabolisable coefficients of dry matter, crude protein (CP), neutral detergent fibre (NDF), and gross energy (GE). There was no interaction between the energetic reductions and inclusion of xylanases on performance variables of the birds. The 70 kcal/kg reduction led to better results for feed intake, weight gain, and feed conversion, while the 140 kcal/kg reduction led to worse results. In terms of jejunum morphometry, there was no interaction between energy reductions and inclusion of xylanases, although xylanases increased villi height and villus:crypt ratio. There was an interaction between energy reduction and inclusion of enzymes for the metabolisability coefficients of CP, NDF, and GE, leading to improvements in these coefficients. Xylanases A and B are effective in energy-reduced corn and soybean meal diets for 15 to 35-day-old quail.(AU)

Glucose homeostasis in the euryhaline crab Cytograpsus angulatus: Effects of the salinity in the amylase, maltase and sucrase activities in the hepatopancreas and in the carbohydrate reserves in different tissues.

We studied the existence, biochemical characteristics and response to different environmental salinities of amylase, maltase and sucrase activity in the intertidal euryhaline crab Cyrtograpsus angulatus (Dana, 1852) along with the response to distinct salinities of glycogen and free glucose content in storage organs. Amylase, maltase and sucrase activities were kept over a broad range of pH and temperature and exhibited Michaelis-Menten kinetics. Zymography showed the existence of two amylase forms in crabs exposed to 35 (osmoconformation) and low (6-10psu; hyper-regulation) or high (40psu) (hypo-regulation) salinities. Carbohydrases activity in the hepatopancreas and glycemia were not affected in crab exposed to different environmental salinities. In 6 and 40psu, the glycogen content in anterior gills was lower than in 35psu. In 6, 10 and 40psu, glycogen concentration in hepatopancreas, muscle and posterior gills were similar to that in 35psu. Free glucose concentration in chela muscle was higher in 6 and 40psu than in 35psu. The existence and biochemical characteristics of carbohydrases activity and the adjustments in concentration of glycogen in anterior gills and free glucose in chela muscle suggests the ability to perform complete hydrolysis of glycogenic substrates and to keep glucose homeostasis in relation to acclimation to different salinity conditions.

Effects of multi-carbohydrase and phytase on standardized ileal digestibility of amino acids and apparent metabolizable energy in canola meal fed to broiler chicks.

Two assays were conducted to evaluate nutritive value of canola meal (CM) fed to broiler chicks without or with a multi-carbohydrase (MC) preparation (700 U α-galactosidase, 2,200 U galactomannanase, 30,000 U xylanase, and 22,000 U ß-glucanase per kg of diet) and phytase (Phy, 500 FTU per kg of diet). Assay 1 determined apparent total tract digestibility (ATTD) of nutrients and metabolizable energy (AME) by the difference method. Assay 2 determined apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids by the index method. Two reference diets (RD) - an 85% corn based and a 5% casein-cornstarch diet fortified with vitamins and minerals - were made for assays 1 and 2, respectively. For each assay, the test diets were made by mixing RD and CM 7:3 wt/wt basis and fed without or with MC or Phy or combination. A total of 245 day-old male broilers (Cobb 500) was allocated to 5 treatments to give 7 replicates (7 birds/cage). The birds were fed a commercial diet from day zero to 10 followed by assay 1 fed from d 11 to 18 and assay 2 fed from d 19 to 21. Excreta samples were collected on d 15 to18, and all birds were slaughtered on d 21 for ileal digesta. There was an interaction (P < 0.05) between MC and Phy on ATTD of DM, N, and P. There was no interaction (P > 0.05) between MC and Phy on AMEn; however, MC and Phy individually improved AMEn retention. Enzymes interacted (P < 0.05) on SID of Arg, His, Leu, Met, Thr, Ala, Asp, Gln, and Gly. In this context, feeding a combination of MC and Phy resulted in higher (P < 0.05) SID of Arg, His, Met, and Thr relative to single activity or control. Both enzymes improved (P < 0.05) SID of Lys independently. The combination of carbohydrase and Phy may be an effective strategy to improve amino acid utilization in CM for poultry.

Energetic values and performace of broilers feeding sorghum and soybean meal based diets supplemented with B-glucanase and B-xylanase / Valores energéticos e desempenho de frangos de corte alimentados com dietas a base de sorgo e farelo de soja suplementadas com B-glucanase and B-xylanase

Grains, brans, and vegetable meals may contain non-starch polysaccharides (NSP), which increases viscosity in the gastrointestinal tract (GIT) and interfere with the digestion and absorption of nutrients. This study aimed to evaluate the performance and determine the metabolizable energy of a sorghum-based broiler diet with and without the supplementation of an enzymatic complex. The experiments were conducted in a completely randomized design with 1200 chickens, using sorghum-based feed with and without the addition of 50 g of enzyme-CCE complex (β-glucanase and β-xylanase), and with two levels of metabolizable energy (ME kg-1): ME; ME + CCE; reduced ME (-50 kcal kg-1); and reduced ME + CCE. The data were subjected to an analysis of variance and the means were compared using a Tukeys test at the 5% significance level. At 42 and 47 days of age, the living weight of the birds fed with the reduced ME was low, while birds fed with reduced ME + CCE had the same weight as those fed with other energy diets (ME and ME + CCE). Feed conversion was poorest at 47 days of age for the birds on reduced ME diet. In the metabolic test (with fattening diets) to determine AME and AMEn, the reduced ME diet had the lowest result, confirming the effect of the addition of enzymes. The addition of CCE to sorghum-based diets provides enough enzymatic activity to increase the metabolizable energy of the diet (50 kcal of AME) and influence the growth performance of broilers at the slaughtering age.(AU)

Grãos, farelos e farinhas de origem vegetal podem conter polissacarídeos não amiláceos (PNAs), que aumentam a viscosidade do trato grastro intestinal (TGI) e interferem na digestão e absorção de nutrientes. Objetivou-se avaliar o desempenho e determinar à energia metabolizável da dieta a base de sorgo, com e sem adição do complexo enzimático (CCE), para frangos de corte. Foram utilizados 1200 frangos de corte, distribuídos em delineamento inteiramente casualisado, alimentados com dois níveis de Energia Metabolizável (EM kg-1), em dietas à base de sorgo com ou sem a adição de 50g de completo enzimático (β-glucanase e β-xylanase): Energia Metabolizável (EM); Energia Metabolizável com Complexo Enzimático (EM + CCE); EM reduzida (-50kcal EM kg-1) e EM reduzida + CCE. Os dados obtidos foram submetidos à análise de variância e as médias dos tratamentos em cada variável estudada foram comparadas entre si pelo Teste de Tukey a nível de 5% de significância. Aos 42 e 47 dias de idade o peso vivo das aves alimentadas com EM reduzida foi menor, enquanto que EM reduzida + CCE apresentou peso igual ao das dietas EM e EM + CCE. A conversão alimentar aos 47 dias de idade na dieta EM reduzida apresentou o pior resultado. No teste metabólico, com as dietas de engorda, os valores de Energia Metabolizável Aparente (EMA) e Energia Metabolizável Aparente corrigida para nitrogênio (EMAn) foram menores na dieta com EM reduzida, comprovando o efeito da adição das enzimas. Concluiu-se que a adição de complexo enzimático (β-glucanase and β-xylanase) melhora o desempenho de frangos de corte e aumenta a energia metabolizável aparente da dieta em 50 kcal.(AU)

Energetic values and performace of broilers feeding sorghum and soybean meal based diets supplemented with B-glucanase and B-xylanase / Valores energéticos e desempenho de frangos de corte alimentados com dietas a base de sorgo e farelo de soja suplementadas com B-glucanase and B-xylanase

Grains, brans, and vegetable meals may contain non-starch polysaccharides (NSP), which increases viscosity in the gastrointestinal tract (GIT) and interfere with the digestion and absorption of nutrients. This study aimed to evaluate the performance and determine the metabolizable energy of a sorghum-based broiler diet with and without the supplementation of an enzymatic complex. The experiments were conducted in a completely randomized design with 1200 chickens, using sorghum-based feed with and without the addition of 50 g of enzyme-CCE complex (β-glucanase and β-xylanase), and with two levels of metabolizable energy (ME kg-1): ME; ME + CCE; reduced ME (-50 kcal kg-1); and reduced ME + CCE. The data were subjected to an analysis of variance and the means were compared using a Tukey’s test at the 5% significance level. At 42 and 47 days of age, the living weight of the birds fed with the reduced ME was low, while birds fed with reduced ME + CCE had the same weight as those fed with other energy diets (ME and ME + CCE). Feed conversion was poorest at 47 days of age for the birds on reduced ME diet. In the metabolic test (with fattening diets) to determine AME and AMEn, the reduced ME diet had the lowest result, confirming the effect of the addition of enzymes. The addition of CCE to sorghum-based diets provides enough enzymatic activity to increase the metabolizable energy of the diet (50 kcal of AME) and influence the growth performance of broilers at the slaughtering age.

Grãos, farelos e farinhas de origem vegetal podem conter polissacarídeos não amiláceos (PNAs), que aumentam a viscosidade do trato grastro intestinal (TGI) e interferem na digestão e absorção de nutrientes. Objetivou-se avaliar o desempenho e determinar à energia metabolizável da dieta a base de sorgo, com e sem adição do complexo enzimático (CCE), para frangos de corte. Foram utilizados 1200 frangos de corte, distribuídos em delineamento inteiramente casualisado, alimentados com dois níveis de Energia Metabolizável (EM kg-1), em dietas à base de sorgo com ou sem a adição de 50g de completo enzimático (β-glucanase e β-xylanase): Energia Metabolizável (EM); Energia Metabolizável com Complexo Enzimático (EM + CCE); EM reduzida (-50kcal EM kg-1) e EM reduzida + CCE. Os dados obtidos foram submetidos à análise de variância e as médias dos tratamentos em cada variável estudada foram comparadas entre si pelo Teste de Tukey a nível de 5% de significância. Aos 42 e 47 dias de idade o peso vivo das aves alimentadas com EM reduzida foi menor, enquanto que EM reduzida + CCE apresentou peso igual ao das dietas EM e EM + CCE. A conversão alimentar aos 47 dias de idade na dieta EM reduzida apresentou o pior resultado. No teste metabólico, com as dietas de engorda, os valores de Energia Metabolizável Aparente (EMA) e Energia Metabolizável Aparente corrigida para nitrogênio (EMAn) foram menores na dieta com EM reduzida, comprovando o efeito da adição das enzimas. Concluiu-se que a adição de complexo enzimático (β-glucanase and β-xylanase) melhora o desempenho de frangos de corte e aumenta a energia metabolizável aparente da dieta em 50 kcal.

Inhibitory effects of an extract of fruits of Physalis peruviana on some intestinal carbohydrases / Evaluación in vitro del efecto de un extracto de frutos de Physalis peruviana sobre algunas carbohidrasas intestinales

Physalis peruviana is an Andean specie whose fruits are eaten as food and also has been reported in Colombian folk medicine for diabetes mellitus treatment. In addition, previous pharmacological studies on diabetic Wistar rats, an extract of P. peruviana fruits has been showed antidiabetic activity. In order to deepen in P. peruviana action mode as antidiabetic, in this investigation were determinate the Inhibitory Concentrations 50 (IC50) of P. peruviana fruits crude extract on alpha glucosidase from S. cerevisiae and from intestinal rat powder, on maltase and alpha amylase enzymes. The kinetic behavior of the extract on each enzyme was also investigated, and the enzyme constant (Km) and maximum rate (Vmax) were determined. Extract of fruits of P. peruviana showed different IC50 for alpha glucosidase from S. cerevisiae and intestinal rat powder, suggesting greater affinity for the enzyme of mammalian source (4114.7 and 3552.7 µg/mL, respectively). For maltase, the IC50 was close to that obtained for alpha glucosidase (4191.0 µg/mL) while for alpha amylase, the extract exhibited the highest inhibition (IC50: 619.9 g/mL). Regarding kinetic behavior, the extract showed competitive inhibition on alpha-glucosidase and maltase, and on the non-competitive type of alpha amylase. These suggest that inhibition of intestinal carbohydrases is one of the modes of action for the antidiabetic activity of fruits of P. peruviana.

Physalis peruviana es una especie andina, cuyos frutos además de ser usados como alimento, son empleados en la medicina tradicional para el tratamiento de la diabetes mellitus. Además, estudios farmacológicos previos en ratas Wistar han demostrado actividad antidiabética de extractos de frutos de P. peruviana. Con el fin de profundizar en el modo de acción de la actividad antidiabética de los frutos de P. peruviana, en la presente investigación se determinó la concentración inhibitoria 50 (CI50) del extracto crudo de frutos de P. peruviana, sobre las enzimas alfa glucosidasa obtenida de S. cerevisiae y de polvo intestinal de rata, maltasa y alfa amilasa. El comportamiento cinético del extracto sobre cada una de las enzimas también fue investigado y la constante enzimática (Km) y la velocidad máxima (Vmax) fueron determinadas. El extracto de frutos de P. peruviana, mostró diferentes valores de CI50 para alfa glucosidasa obtenida de S. cerevisiae y para la obtenida de polvo intestinal de rata, sugiriendo una mayor afinidad por la enzima de origen mamífero (4114,7 and 3552,7 µg/mL, respectivamente). Para maltasa la CI50 fue cercana a la obtenida para alfa glucosidasa (4191,0 µg/mL), mientras para alfa amilasa, el extracto presentó la mayor inhibición (CI50: 619,9 g/mL). Respecto al comportamiento cinético, el extracto mostró inhibición de tipo competitiva sobre alfa glucosidasa y maltasa y no competitiva sobre alfa amilasa. Los resultados sugieren que la inhibición de carbohidrasas intestinales es uno de los modos de acción de los frutos de P. peruviana como agente antidiabético.

Efficacy and Metabolizable Energy Equivalence of an -Amylase--Glucanase Complex for Broilers

A trial was conducted to evaluate the effects of adding an exogenous -amylase--glucanase complex produced from Bacillus amyloliquefaciens on the growth performance, carcass yield, and relative AME bioequivalence in broilers fed corn-soy diets from 1 to 40 d of age. One thousand seven hundred and fifty one-day-old Cobb x Cobb 500 slow-feathering male broilers were randomly allotted to seven treatments with 10 replicates of 25 birds each as follows: control diet (C); C diet with reduction of 60 (C-60), 90 (C-90), or 120 (C-120) kcal AME/kg; C diet with reduction of 120 kcal AME/kg and supplemented with 200 (C-120-200), 300 (C-120-300), or 400 (C-120-400) mg of the enzyme complex/kg. Each g of the enzyme complex corresponded to 200 kilo-Novo -amylase and 350 fungal -glucanase units. On d 40, eight birds were randomly taken from each pen and processed to evaluate carcass and commercial cuts yields. Percent mortality was not affected by the treatments (p > 0.05). Live performance, as indicated by BW gain (BWG) linearly decreased (p 0.05) and FCR linearly increased with the reduction in AME. Birds fed diets supplemented with the enzyme complex showed weekly improvements in BWG and FCR. There were no effects of the treatments on the yield of the carcass or of commercial cuts; however, abdominal fat was significantly lower (p 0.0343) in birds fed the C-120-400 compared to the C-120 feeding program (1.67% vs. 1.90%); all other treatments were intermediate. Average AME equivalence of the enzyme complex varied weekly. Estimations for the entire period were 40, 46, and 56 kcal for BWG and 58, 76, and 99 kcal AME/kg for FCR (p 0.001) for the diets containing 200, 300, and 400 mg enzyme complex/kg, respectively.(AU)