Fig 1 - uploaded by Juan Tricarico
Content may be subject to copyright.
Growth (optical density at 660 nm) on soluble potato starch (1 g/l) in the absence (empty square) or presence (solid square) of supplemental-amylase (0.06 DU/ml) by (A) Butyrivibrio fibrisolvens D1; (B) Selenomonas ruminantium GA192; (C) Megasphaera elsdenii T81; (D) Streptococcus bovis S1; (E) Butyrivibrio fibrisolvens 49; and (F) Butyrivibrio fibrisolvens A38.
Source publication
Research in the area of dietary enzyme supplements for ruminant diets has primarily focused on fibrolytic enzymes, while activities involved in the process of starch digestion have been largely ignored. Since starch represents a major component in diets fed to highly productive cattle, the use of enzymatic dietary supplements to manipulate starch d...
Contexts in source publication
Context 1
... Microbial growth was estimated in each culture by measuring optical density at 600 nm over time. Each experi- ment consisted of either two or three replicates per treatment. As expected, Streptococcus bovis S1 and Butyrivibrio fibrisolvens 49 grew rapidly on starch-containing medium and -amylase supplementation had no effects on their growth (Fig. 1). Butyrivibrio fibrisol- vens A38 grows equally well on maltose and starch (Cotta, 1988) and grew more slowly in the presence of supplemental -amylase in this experiment. Conversely, Butyrivibrio fibri- solvens D1, Selenomonas ruminantium GA192 and Megasphaera elsdenii T81 only grew poorly or not at all on starch. However, these ...
Context 2
... grew more slowly in the presence of supplemental -amylase in this experiment. Conversely, Butyrivibrio fibri- solvens D1, Selenomonas ruminantium GA192 and Megasphaera elsdenii T81 only grew poorly or not at all on starch. However, these non-amylolytic species grew rapidly when supplemental -amylase was included in the starch-containing medium (Fig. ...
Citations
... Additionally, exogenous amylolytic enzymes have been evaluated to optimize starch utilization and increase animal performance in feedlot diets (Tricarico et al., 2007;DiLorenzo et al., 2011). However, amylolytic enzymes do not seem to increase ruminal starch digestion but induce shifts in the molar proportion of butyrate and acetate by exposing oligosaccharides for cross-feeding by rumen microbes (Tricarico et al., 2008). Some studies have also reported an increase in DM and NDF digestibility (DiLorenzo et al., 2011;Vera et al., 2012) leading to changes in ruminal fermentation parameters , while others found no effects (He et al., 2014;He et al., 2015). ...
... Additionally, the rise in fibre-degrading bacteria may help improve fibre digestion, contributing to enhanced fibre passage rates. Tricarico et al. (2008) reported improved performance traits in calves, steers, and heifers on a highconcentrate diet supplemented with prebiotic feeds based on S. cerevisiae and/or Aspergillus oryzae. Additionally, Geng et al. (2016) added to feed of finishing bulls a probiotic or a synbiotic based on S. cerevisiae. ...
... These results are similar to the research of Tricarico et al. (2005), who found improved acetate and butyrate as well as reduced propionate molar proportions in steers and lactating dairy cows with dietary addition of α-amylase. In a subsequent study, Tricarico et al. (2008) hypothesized a cross-feeding mechanism that the supplemented α-amylase and fibrolytic enzymes would hydrolyze amylose, cellulose and xylans into oligosaccharides, thereby providing substrate to nonfibrolytic or non-amylolytic bacteria, giving these bacteria a competitive advantage. This hypothesis could probably explain the current study. ...
This research communication reports the effects of a compound enzyme preparation consisting of fibrolytic (cellulase 3500 CU/g, xylanase 2000 XU/g, β-glucanase 17 500 GU/g) and amylolytic (amylase 37 000 AU/g) enzymes on nutrient intake, rumen fermentation, serum parameters and production performance in primiparous early-lactation (47 ± 2 d) dairy cows. Twenty Holstein–Friesian cows in similar body condition scores were randomly divided into control (CON, n = 10) and experimental (EXP, n = 10) groups in a completely randomized single-factor design. CON was fed a basal total mixed ration diet and EXP was dietary supplemented with compound enzyme preparation at 70 g/cow/d. The experiment lasted 4 weeks, with 3 weeks for adaptation and then 1 week for measurement. Enzyme supplementation significantly increased diet non-fibrous carbohydrates (NFC) content as well as dry matter intake (DMI) and NFC intake ( P < 0.05). EXP had increased ruminal butyrate and isobutyrate percentages ( P < 0.01) but decreased propionate and valerate percentages ( P < 0.05), as well as increased serum alkaline phosphatase activity and albumin concentration ( P ≤ 0.01). Additionally, EXP had increased milk yield (0.97 kg/d), 4% fat corrected milk yield and energy corrected milk yield, as well as milk fat and protein yield ( P < 0.01). In conclusion, dietary supplementation with a fibrolytic and amylolytic compound enzyme preparation increased diet NFC content, DMI and NFC intake, affected rumen fermentation by increasing butyrate proportion at the expense of propionate, and enhanced milk performance in primiparous early-lactation dairy cows.
... Pérez et al. (2009) stated that amylolytic enzymes can produce as much as 600 metabolizable glucose units by targeting α-glycosidic bonds and increasing starch digestion (Rojo et al. 2007). Tricarico et al. (2008) hypothesized that amylolytic enzymes work by cross-feeding mechanisms of ruminal bacteria through oligosaccharides produced by the enzymes, creating modified products of ruminal fermentation. Further, supplementation of amylolytic enzymes to ruminants showed mixed results. ...
Competition among animal feeds, human foods, and demands for biofuel production forces livestock producers to feed their animals on the available low-cost raw materials. Ruminant farmers provide their animals forage-based diets to reduce the cost of feeding and to keep normal rumen function. However, only forage feeding does not sometimes provide animals enough available energy and protein required for milk production and growth. The major problem with feeding low-quality forages or raw materials is their content of rigid and less-digestible cell wall components. However, ruminants have a digestive system with a complex and dynamic microbiota and microbial activities that allow them to utilize plant cell walls and degrade them to be utilized as energy sources by host. The ability of ruminants to degrade structural polysaccharides (e.g., cellulose, hemicellulose, and lignin) has a limit, and under perfect conditions, is less than 65%. Different approaches have been developed to improve nutrient digestibility and production responses in ruminants including chemical, physical, and biological strategies. Exogenous enzymes have gained an increasing interest to improve the nutritive value of ruminant feeds. The aim of administration of exogenous enzymes is to improve fiber degradation and efficiency of feed utilization by enhanced nutrient utilization and reduce the wastage. Exogenous enzymes are capable of breaking down specific bonds in carbohydrates, starch, protein, fats, cellulose, hemicellulose, pectin, glycoproteins, and lignin. Lactobacillus acidophilus, L. plantarum, Streptococcus faecium, and Bacillus subtilis are the main bacterial species that produce a wide array of exogenous enzymes, while Trichoderma reesei, Aspergillus oryzae, and Saccharomyces cerevisiae are the main fungal enzyme producers. The ability of exogenous enzymes to improve growth performance in feedlot cattle and small ruminants as well as milk production in dairy cattle has been demonstrated, but with inconsistent effects. Enzyme type and preparation, application method, level of enzyme supplementation, diets, animals and interaction between enzymes and feeds are the factors that determine the response to enzyme supplementation and are responsible for variations among experiments. This chapter reviews information about sources of enzymes, enzyme activity, modes of action, optimal inclusion, and response of ruminant to dietary applications of various enzymes.
... A possible explanation for the increase in amylase occurring after simulated digestion in F1F and not in F8F is the difference in the concentration of carbohydrates and complex polysaccharides being more significant in the second formulation, which may have required more excellent enzymatic action and, consequently, consumption. Amylase can modify the availability of the metabolic profile of starch digestion, bringing advantages to the fermentation process (Gallo et al. 2016;Tricarico et al. 2008). In vitro, gastrointestinal digestion showed reduced lactic acid bacteria and yeast and increased acetic acid bacteria in the liquid phase for both formulations ( Table 6). ...
This work aimed to study and characterize a product based on vegetable extract of quinoa (WVEQ) fermented with water kefir grains. The effect of sucrose concentration (SC), inulin concentration (IC), and xanthan gum (XG) concentration were evaluated using a central composite design (CCD) 2³. They were subsequently characterized regarding cellular growth of the grains, beverage yield, pH, soluble solids, carbon dioxide (CO2) production, lactic acid, and ethanol production. Therefore, for the final stage, two formulations (F1 and F8) of the CCD were chosen to be characterized in terms of proximate composition, microbiological composition of the kefir culture, analysis of organic compounds, sensory analysis, and enzymatic and microbiological characterization before and after simulation of in vitro gastrointestinal digestion. In the two chosen products, one can see that fermentation optimized the bioavailability of proteins due to the high proteolytic activity of the microorganisms in kefir and the increase in lipid content. In identifying microorganisms, there was a prevalence of Saccharomyces sp. yeasts. In the sensory analysis, the F8 formulation showed better results than the F1 formulation. In vitro, gastrointestinal digestion showed reduced lactic acid bacteria and yeast and increased acetic acid bacteria in the liquid phase for both formulations. In the enzymatic profile, there was a reduction in all enzymes analyzed for both formulations, except for amylase in F1, which went from 14.05 U/mL to 39.41 U/mL. Therefore, it is concluded that using WVEQ as a substrate for the product appears to be a viable alternative with nutritional and technological advantages for serving a specific market niche.
... It was reported that supplementing α-amylase (AM) in diets increased average daily gain (ADG) in finishing beef (3) and improved lactation performance in dairy cows (4,5). Moreover, some studies found that AM addition increased rumen butyrate molar proportion (6), stimulated B. fibrisolvens D1 growth in vitro (7), and increased AM activity and rumen propionate molar proportion of cows (8). Others reported that supplementation with AM increased total-tract digestibility of organic matter (OM), dry matter (DM), neutral detergent fiber (NDF), and crude protein (CP) in cows (4,9). ...
... Moreover, the increase in AM activity was mainly caused by the positive responses of amylolytic bacteria, showing a synergistic effect of exogenous AM and rumen microbes, as reported by Noziere et al. (8). Similarly, other studies indicated that AM supplementation increased ruminal propionate molar proportion and AM activity in dairy cows (8) and stimulated B. fibrisolvens D1 growth in vitro (7). However, according to Tricarico et al. (6), AM addition increased the butyrate and acetate molar proportions and the acetate-to-propionate ratio. ...
This study evaluated the impacts of α-amylase (AM) and coated α-amylase (CAM) on bull performance, nutrient digestibility, and ruminal fermentation. This study randomized 60 Holstein bulls of 365 ± 11.5 days of age and 457.5 ± 9.35 kg body weight into three groups: without AM addition, adding AM 0.6 g/kg dry matter (DM), and adding CAM 0.6 g AM/kg DM, separately. This whole experimental period was 80 days, including a 20-day adaptation period and a 60-day data and sample acquisition period. In comparison with the unsupplemented control, dry matter intake (DMI) was unaltered; however, average daily gain (ADG) and feed efficiency (FE) were greater for AM or CAM addition. Bulls receiving AM or CAM supply had greater total-tract nutrient digestibility, ruminal total volatile fatty acids (VFA) content, propionate molar proportion, cellulolytic enzyme and AM activities, and the number of microorganisms. In addition, the activities of AM and trypsin in the jejunum and ileum and glucose, albumin, and total protein concentrations in serum were greater for AM or CAM addition compared to the control. When comparing the supplementation mode of AM, bulls receiving CAM addition had greater ADG and FE. The crude protein and starch digestibility and intestinal AM and trypsin activity were higher, while acid detergent fiber (ADF) digestibility was lower for CAM addition than for AM addition. The lower propionate molar proportion and cellobiase and carboxymethyl cellulase activities, together with Ruminococcus albus, Ruminococcus flavefaciens, and Fibrobacter succinogenes populations were observed for CAM addition compared with AM addition. However, there were greater glucose, albumin, and total protein concentrations in serum after adding CAM. According to the data, the supply of AM improved ADG, nutrient digestion, and rumen fermentation. Notably, the optimum supplementation mode was in the form of CAM in bulls.
... Researchers found that the inclusion of amylase promotes rapid release of starch oligosaccharides that are used by both amylolytic and non-amylolytic bacteria [34], for this reason starch is fermented quickly. This was observed in our study, in both groups of steers that consumed amylase in the diet, the starch digestibility coefficient was higher; as well as increasing the concentration of total volatile fatty acids in the rumen; but this was not enough when the exogenous enzyme was added to the diet to enhance weight gain. ...
The objective of this study was to evaluate if the inclusion of a blend composed of exogenous enzymes (amylase, protease, cellulase, xylanase and beta glucanase) in the individual and combined form in the feedlot steers diet has benefits on the physiology, rumen fermentation, digestibility and fatty acid profile in rumen and meat. The experiment used 24 animals, divided into 4 treatments, described as: T1-CON, T2-BLEND (0.5 g mixture of enzyme), T3-AMIL (0.5 g alpha-amylase), T4-BLEND+AMIL (0.5 g enzyme blend+ 0.5 g amylase). The concentration of mineral matter was higher in the meat of cattle of T4-BLEND+AMIL. A higher proportion of monounsaturated fatty acids was observed in the T3-AMIL group when compared to the others. The percentage of polyunsaturated fatty acids was higher in the T2-BLEND and T4-BLEND+AMIL compared to the T1-CON. The combination of exogenous enzymes in the diet positively modulate nutritional biomarkers, in addition to benefits in the lipid and oxidative profile meat.
... Tal capacidade pode ser comprovada em estudo com adição de culturas vivas de Aspergillus oryzae, Saccharomyces cerevisiae e de seus respectivos extratos, utilizados como suplementos alimentares na dieta de ruminantes. Pesquisas demonstram que esses aditivos microbianos podem melhorar o ganho de peso, a digestibilidade total da fibra (TRICARICO et al., 2008) Nessa análise, foi possível observar que, mesmo dentro de um único gênero, como o Aspergillus, houve variações individuais entre os valores de AC dos 46 isolados avaliados. ...
O ambiente ruminal é composto por inúmeros microrganismos, onde cada um deles possui um papel fundamental no aproveitamento das dietas ingeridas pelos ruminantes, os fungos são seres unicelurares que atuam degradando as porções fibrosas dessa dieta. Objetivou-se neste estudo avaliar a atividade celulolítica (AC) de fungos micelianos aeróbios provenientes do trato digestório de ovinos. Foram identificados e avaliados 38 isolados de fungos micelianos provenientes de borregos e ovelhas, após o reisolamento, amostras desses microrganismos foram inoculadas em meio C, contendo celulose microcristalina a 1% como única fonte de carbono e incubadas em estufa a 37 °C, em triplicata, por um período de três dias. As leituras da AC foram realizadas após 24, 48 e 72 horas de incubação. Foi adicionado solução contendo vermelho congo e após 15 minutos, as placas foram lavadas com 15 mL de solução 1M de NaCl. Os índices de AC foram calculados pela razão entre o halo de degradação de celulose e o halo da colônia. Dois isolados do gênero Aspergillus e um Trichoderma sp. apresentaram índice AC maior que um, apresentando bom crescimento ao utilizarem esse polissacarídeo com única fonte de carbono. Futuros estudos devem avaliar o potencial desses fungos como probióticos na alimentação de ruminantes ou para microbiologia industrial. Palavras-chave: Ovinos. Celulose. Trato digestório. Norte de Minas Gerais. Abstract The ruminal environment is composed of numerous microorganisms, each of which plays a fundamental role in the use of the diets ingested by ruminants, fungi are unicellular beings that act by degrading the fibrous portions of this diet. The objective of this study was to evaluate the cellulolytic activity (CA) of aerobic mycelial fungi from the digestive tract of sheep. Thirty-eight isolates of mycelial fungi from lambs and sheep were identified and evaluated, after re-isolation, samples of these microorganisms were inoculated in medium C, containing 1% microcrystalline cellulose as the only carbon source, and incubated in an oven at 37 °C, in triplicate, for a period of three days. AC readings were performed after 24, 48 and 72 hours of incubation. Solution containing congo red was added and after 15 minutes, the plates were washed with 15 mL of 1M NaCl solution. CA indices were calculated by the ratio between the cellulose degradation halo and the colony halo. Two isolates of the genus Aspergillus and one Trichoderma sp. showed CA index greater than one, showing good growth when using this polysaccharide as a single carbon source. Future studies should assess the potential of these fungi as probiotics in ruminant feed or for industrial microbiology. Keywords: Sheep. Cellulose. Digestive tract. North of Minas Gerais.
... The amylolytic enzyme used in this experiment (AMAIZE TM , Alltech Inc., Nicholasville, KY, USA) was powdered Aspergillus oryzae extract, which mainly contains α-amylase or 1,4-α-D-glucan glucanohydrolase (Tricarico et al., 2008) and is resistant to degradation by rumen microbes (Klingerman et al., 2009). One α-amylase dextrinizing unit (DU) is defined as the quantity of enzyme required to dextrinize soluble starch at the rate of 1 g h -1 at 30ºC and pH 4.8. ...
... One α-amylase dextrinizing unit (DU) is defined as the quantity of enzyme required to dextrinize soluble starch at the rate of 1 g h -1 at 30ºC and pH 4.8. The α-amylase concentration in the A. oryzae-based supplement was 600 DU g -1 (Tricarico et al., 2008). ...
... Increases in nutrient digestibility may be due to the synergism between endogenous and exogenous enzymes, increasing overall ruminal enzymatic activity and hydrolysis (Meale et al., 2014). On the other hand, when α-amylase of A. oryzae is used, the overall nutrient digestibility increases since the maltodextrins produced from the hydrolysis of native starch can be used by a wide variety of ruminal bacteria, including amylolytic and non-amylolytic species (oligosaccharide cross-feeding hypothesis) (Tricarico et al., 2008). The quadratic response in nutrient digestibility can be explained by the abovementioned oligosaccharide cross-feeding hypothesis, in which low exogenous enzyme concentrations would not produce sufficient oligosaccharides for effective cross-feeding, whereas high enzyme doses, or prolonged exposure to enzymes, would extensively hydrolyze polymers to di-and monosaccharides, thereby failing to support an effective cross-feeding mechanism (Tricarico et al., 2008). ...
Highlights Increased levels of amylolytic enzymes in the diet did not change ewe's comsumption. Changes in nutrients digestibility were observed. Inclusion of exogenous amylolytic enzyme can improve dry matter digestibility. Abstract The aim was to evaluate if the inclusion of exogenous amylolytic enzyme affect the nutrient intake and digestibility in ewes fed high-concentrate diets containing flint corn. Five Santa Inês × Dorper crossbred ewes (54.04 ± 4.5 kg and aged 8 months) were used in a 5 x 5 Latin square design. All animals were housed in individual metabolic cages for 60 days. The treatments consisted of a control diet (without amylolytic enzyme) and four inclusion levels of an amylolytic enzyme (3,000, 6,000, 9,000, and 12,000 α-amylase dextrinizing units [DU] kg-1 dry matter [DM]). The enzyme was mixed into the feed at the time of supply to the animals. Data were analyzed by ANOVA, and orthogonal polynomial contrasts were used. Nutrient intake was not influenced by amylolytic enzyme inclusion. The digestibility of DM, organic matter, neutral detergent fiber, total carbohydrates, non-fibrous carbohydrates, and gross energy showed a quadratic increase with enzyme inclusion (P<0.05), with maximum values at levels of 7,600, 7,500, 6,300, 7,500, 7,400, and 7,800 DU kg-1 DM, respectively. Total digestible nutrients of diets also showed a quadratic increase, with a maximum value of 894 g kg-1 at a level of α-amylase activity of 7,786 DU kg-1 DM. The inclusion of the exogenous amylolytic enzyme from 6,300 to 7,800 DU kg-1 DM doesn't alter nutrient intake and improves the digestibility in ewes fed high-concentrate diets. Resumo Objetivou-se avaliar se a inclusão de enzima amilolítica exógena afeta a ingestão e a digestibilidade dos nutrientes em ovelhas alimentadas com dietas de alto teor de concentrado contendo milho flint. Foram usadas cinco ovelhas mestiças Santa Inês × Dorper (54,04 ± 4,5 kg e 8 meses de idade) em um quadrado latino 5x5. Todos os animais foram alojados em gaiolas metabólicas individuais por 60 dias. Os tratamentos consistiram em uma dieta controle (sem enzima amilolítica) e quatro níveis de enzima amilolítica (3.000; 6.000; 9.000 e 12.000 unidades de dextrinização [UD] de α-amilase kg-1 de matéria seca [MS]). A enzima foi misturada com o alimento no momento do fornecimento aos animais. Os dados foram analisados por ANOVA e contrates ortogonais polinomiais. O consumo de nutrientes não foi influenciado pela inclusão da enzima amilolítica. A digestibilidade da MS, matéria orgânica, fibra em detergente neutro, carboidratos totais, carboidratos não fibrosos e energia bruta apresentaram efeito quadrático com a inclusão de enzima (P<0,05), com valores máximos nos níveis de inclusão da enzima 7.600; 7.500; 6.300; 7.500; 7.400 e 7.800 UD kg-1 MS, respectivamente. Os nutrientes digestíveis totais também apresentaram efeito quadrático, com máximo valor de 894 g kg-1 no nível de atividade de α-amilase de 7.786 UD kg-1 MS. A inclusão da enzima amilolítica exógena entre 6.300 e 7.800 UD kg-1 MS não modifica o consumo de nutrientes e melhora a digestibilidade de ovelhas alimentadas com dietas de alto teor de concentrado. Palavras-chave: Alfa-amilase. Amido. Nutrição. Ovelhas.
... The amylolytic enzyme used in this experiment (AMAIZE TM , Alltech Inc., Nicholasville, KY, USA) was powdered Aspergillus oryzae extract, which mainly contains α-amylase or 1,4-α-D-glucan glucanohydrolase (Tricarico et al., 2008) and is resistant to degradation by rumen microbes (Klingerman et al., 2009). One α-amylase dextrinizing unit (DU) is defined as the quantity of enzyme required to dextrinize soluble starch at the rate of 1 g h -1 at 30ºC and pH 4.8. ...
... One α-amylase dextrinizing unit (DU) is defined as the quantity of enzyme required to dextrinize soluble starch at the rate of 1 g h -1 at 30ºC and pH 4.8. The α-amylase concentration in the A. oryzae-based supplement was 600 DU g -1 (Tricarico et al., 2008). ...
... Increases in nutrient digestibility may be due to the synergism between endogenous and exogenous enzymes, increasing overall ruminal enzymatic activity and hydrolysis (Meale et al., 2014). On the other hand, when α-amylase of A. oryzae is used, the overall nutrient digestibility increases since the maltodextrins produced from the hydrolysis of native starch can be used by a wide variety of ruminal bacteria, including amylolytic and non-amylolytic species (oligosaccharide cross-feeding hypothesis) (Tricarico et al., 2008). The quadratic response in nutrient digestibility can be explained by the abovementioned oligosaccharide cross-feeding hypothesis, in which low exogenous enzyme concentrations would not produce sufficient oligosaccharides for effective cross-feeding, whereas high enzyme doses, or prolonged exposure to enzymes, would extensively hydrolyze polymers to di-and monosaccharides, thereby failing to support an effective cross-feeding mechanism (Tricarico et al., 2008). ...
The aim was to evaluate if the inclusion of exogenous amylolytic enzyme affect the nutrient intake and digestibility in ewes fed high-concentrate diets containing flint corn. Five Santa Inês × Dorper crossbred ewes (54.04 ± 4.5 kg and aged 8 months) were used in a 5 x 5 Latin square design. All animals were housed in individual metabolic cages for 60 days. The treatments consisted of a control diet (without amylolytic enzyme) and four inclusion levels of an amylolytic enzyme (3,000, 6,000, 9,000, and 12,000 α-amylase dextrinizing units [DU] kg-1 dry matter [DM]). The enzyme was mixed into the feed at the time of supply to the animals. Data were analyzed by ANOVA, and orthogonal polynomial contrasts were used. Nutrient intake was not influenced by amylolytic enzyme inclusion. The digestibility of DM, organic matter, neutral detergent fiber, total carbohydrates, non-fibrous carbohydrates, and gross energy showed a quadratic increase with enzyme inclusion (P<0.05), with maximum values at levels of 7,600, 7,500, 6,300, 7,500, 7,400, and 7,800 DU kg-1 DM, respectively. Total digestible nutrients of diets also showed a quadratic increase, with a maximum value of 894 g kg-1 at a level of α-amylase activity of 7,786 DU kg-1 DM. The inclusion of the exogenous amylolytic enzyme from 6,300 to 7,800 DU kg-1 DM doesn’t alter nutrient intake and improves the digestibility in ewes fed high-concentrate diets.
