Addition Of Amylase Research Articles

Acetic Fermentation of Cagaita Pulp: Technological and Chemical Characteristics

The Brazilian Cerrado region has a rich plant diversity, with fruits that have peculiar and unique sensory characteristics. For these reasons, using these fruits for biotechnological production is a promising alternative, mainly to protect this biome from deforestation and degradation. The production of fermented acetic acid is an option to add value to native fruits and offer the market beverages with better nutritional quality and bioactive compounds. This work aimed to characterize fruits and to develop cagaita (Eugenia dysenterica DC.) acetic fermented beverage. The fruits were subjected to physical-chemical analyses in the first part. Subsequently, different treatments for fermentation were tested using two types of enzymes (amylase and pectinase), two subspecies of Saccharomyces cerevisiae yeast (UFLA CA11 and thermoresistant LNF Angel), and the chaptalization of the must with sucrose (16 °Brix). Alcoholic fermentation was carried out in an incubator with temperature control at 34 ± 1 °C. The pH, total soluble solids, titratable acidity, alcohol content, and density of the fermented products were monitored daily. The chaptalized must with amylase addition and thermoresistant yeast had the best performance during alcoholic fermentation, demonstrating that thermoresistant yeast is an economically advantageous and efficient alternative for the cagaita juice fermentation process. Subsequently, acetic fermentation was carried out using the slow method. Heat-resistant yeast without added enzymes was used to produce cagaita acetic fermented beverages within the parameters of the Brazilian legislation. Furthermore, phenolic compounds and antioxidant activity in the final product were observed. The work demonstrated the possibility of using cagaita fruits in biotechnological processes to produce new food products.

Investigation of Foreign Amylase Adulteration in Honey Distributed in Japan by Rapid and Improved Native PAGE Activity Staining Method.

Foreign amylase addition to honey in an effort to disguise diastase activity has become a widespread form of food fraud. However, since there is no report on the investigation in Japan, we investigated foreign amylases in 67 commercial honeys in Japan. First, the α-glucosidase and diastase activities of honeys were measured, which revealed that only α-glucosidase activity was significantly low in several samples. As both enzymes are secreted from honeybee glands, it is unlikely that only one enzyme was inactivated during processing. Therefore, we suspected the presence of foreign amylase. α-Amylase in honey were assigned using protein analysis software based on LC-QTOF-MS. As a result, α-amylases from Aspergillus and Geobacillus were detected in 13 and 6 out of 67 honeys, respectively. To detect foreign amylases easily, we developed a cost-effective method using native PAGE. Conventional native PAGE failed to separate the α-amylase derived from honeybee and Geobacillus. However, when native PAGE was performed using a gel containing 1 % maltodextrin, the α-amylase from honeybee did not migrated in the gel and the α-amylase could be separated from the other two α-amylases. The results from this method were consistent with those of LC-QTOF-MS method, suggesting that the novel native PAGE method can be used to detect foreign amylases.

The Inhibitory Effects of Amylase and Streptokinase on Minimum Inhibitory Concentration of Antibiotics Used to Treat Gram Negative Bacteria Biofilm Infection on Indwelling Devices.

The study evaluated and compared the effect of adding streptokinase and amylase to antibiotics that are already used in clinical practice to treat Gram negative bacteria biofilm infection on indwelling devices on the antibiotics' minimum inhibitory concentration (MIC). 24h-old biofilms were developed on 96-well plate with eight clinical isolates. MIC of amikacin, cefepime, ceftazidime, colistin, meropenem, and piperacillin-tazobactam, on biofilms were measured before and after the addition of 25U/ml streptokinase and 25μg/ml amylase with microplate reader. The addition of streptokinase reduces the MICs of cefepime, ceftazidime, colistin, meropenem from (16, 16, 8, 4μg/ml) to (8, 1, 1, 0.5μg/ml) in Escherichia coli (isolate 1). While the addition of amylase reduces the MICs of only cefepime, ceftazidime from (16, 16μg/ml) to (2, 4μg/ml) in E. coli (isolate 1). In Pseudomonas aeruginosa (isolate 4), the MICs of amikacin, cefepime, ceftazidime, colistin and meropenem (64, 16, 32, 4, 32μg/ml) reduced to (2, 1, 0.5, 0.25, 0.5μg/ml) with streptokinase and (4, 4, 4, 2, 0.5μg/ml) with amylase respectively. Similar inhibitions were seen in Pseudomonas putida, Proteus mirabilis. We can conclude that the addition of streptokinase and amylase were effective in reducing the MICs of antibiotics that are commonly used to treat Gram negative bacteria biofilm infection on indwelling devices, thereby increasing susceptibility of bacteria to antibiotics. Streptokinase obviously had a greater effect than amylase, implying that it should be prioritized in future in vivo and clinical studies to obtain successful therapy with antibiotics on biofilm infections.

Amilase exógena aumenta a produção de gases e melhora a cinética de digestão ruminal in vitro de grãos de sorgo e milho

ABSTRACT The aim of this study was to evaluate the effect of exogenous amylase on gas production, in vitro dry matter digestibility (IVDMD), and in vitro digestion kinetics of sorghum (Sorghum vulgaris) and two corn hybrids of different grain textures. Ruminal fluid was collected from two rumen-fistulated cows receiving or not exogenous amylase (0.7g kg-1 of dry matter (DM basis)), provided to achieve 396 kilo Novo units kg-1 for amylase activity (DM basis). Gas production was measured after 1, 3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42 e 48 hours of incubation. Amylase increased gas production (mL) by 5.4%. Corn hybrids have higher in vitro dry matter digestibility than sorghum. Exogenous amylase increased the potential of gas production (A) (P=0.01). There was an effect of hybrid for IVDMD (P<0.01). The addition of exogenous amylase increases the in vitro gas production, improves fermentation kinetics, and increases the production of the ammonia nitrogen of corn and sorghum grains, but does not affect in vitro and dry matter digestibility or the short-chain fatty acids production.

The effect of reduction of resistant starch content of faba bean and pea by amylase supplementation on performance, nutrient digestibility, and sialic acid excretion of broiler chickens

The aim of the present study is to investigate the impact of the protein source and amylase addition and the interaction between these two on growth performance and nutrient utilization in particular starch digestibility and AMEN value in broiler chickens. A total of 960 one-day-old male broiler chickens (Ross 308) were utilized in this study. The chickens were randomly classified into four experimental groups, with each group having 30 replications, with eight birds per replication. The groups differed from each other with respect to the source of crude protein in diets (pea or faba bean seeds) and the addition of exogenous amylase (not added or added). The chickens had unlimited access to feed and fresh water and were kept under stable environmental conditions. The body weight and feed intake of the chickens were measured on the 14th and 35th days, with the aim of determining growth performance parameters. At the end of the study, the chickens were sacrificed and samples (excreta and digesta) were collected to estimate the nutritional value. The results showed no interaction (P > 0.05) between the protein source and enzyme addition in any of the analyzed growth performance and nutritional value parameters. The addition of amylase resulted in a higher body weight gain (0–35 days) and feed conversion ratio (each period). Moreover, significantly higher (P < 0.05) dry matter, starch, and amino acid digestibility were observed. Amylase addition resulted in lower excretion of total sialic acids (P < 0.05). The faba bean diet improved the growth performance and had greater nutritional value. In conclusion, amylase addition improved the growth performance and starch and amino acid digestibility. Therefore, it seems that the use of exogenous amylase is effective regardless of the source of protein in broiler chicken nutrition.

The Effect of Exogenous Amylase Supplementation on the Nutritional Value of Pea (Pisum sativum L.) for Broiler Chickens

The present study aimed to investigate whether the exogenous addition of amylase enhances the nutritional value of pea seeds for broiler chickens. In total, 84 1-day-old male broiler chickens (Ross 308) were used for the experimental study. During the first phase of the experiment (1-16 d), all birds in each treatment were fed with a corn-soybean meal reference diet. After this time, the first treatment (control) was still fed the reference diet. In the second and third treatment, 50% of the reference diet was replaced with 50% pea seeds. In addition, the third treatment was supplemented with exogenous amylase. Animal excreta were collected on 21 d and 22 d of the experiment. The birds were sacrificed at the end of the experiment (23 d), and samples of ileum content were collected. The experimental results showed that the exogenous addition of amylase significantly improved (p < 0.05) the apparent ileal digestibility (AID) of the crude protein (CP), starch, and dry matter (DM) of pea. In addition, an improvement in the AID of essential amino acids in pea seeds (except Phe) was observed. The trend in the AMEN values was also noted (p = 0.076). It can be concluded that supplementation with exogenous amylase improves the nutritional value of pea seeds in broiler chicken nutrition.

Improving the Utilization of Food Waste: Conversion of Food Waste into Residual Food Dried Substance and Use of This Material as a Culture Nutrient for Microbial Production of Lactic Acid.

To reduce food waste (FW) disposal costs, many Koreans now convert FW into residual food dried substances (RFDS) using a house-service food drying machine and then dispose of the RFDS. To recycle RFDS, we tested whether RFDS could be used as a culture nutrient to produce value-added microbial chemicals. As a case study, we attempted to produce lactic acid (LA) by cultivating lactic acid bacteria using RFDS. To prepare the culture medium for LA production, we finely ground the RFDS and dissolved it with CaCO3, a pH-controlling agent. Six lactic acid bacteria were tested to improve LA production, with Lactococcus lactis showing the highest LA production. To enhance LA production, three hydrolytic enzymes, amylase, protease, and lipase, were introduced separately or simultaneously into the RFDS medium during the cultivation of the L. lactis strain. The addition of amylase alone was the most effective in increasing LA production. We then investigated the effect of the RFDS concentration on LA production. The highest LA production was achieved when 100g/L of RFDS was used. LA production was scaled up using a 5L bioreactor. During the fermentation, LA production improved to 46.32g/L, which was 1.73-fold higher than that (26.83g/L) obtained from the flask culture. These results show that RFDS from FW can be used as a culture nutrient to produce LA. Our study provides a new and simple FW recycling method and lays the foundation for expanding the usability of FW.

Exploitation of wasted bread as substrate for polyhydroxyalkanoates production through the use of Haloferax mediterranei and seawater.

The use of the halophile microorganism Haloferax mediterranei, able to synthesize poly(hydroxybutyrate-hydroxyvalerate) (PHBV), is considered as a promising tool for the industrial production of bioplastic through bioprocessing. A consistent supplementation of the growth substrate in carbohydrates and minerals is overall necessary to allow its PHBV production. In this work, wasted bread was used as substrate for bioplastic production by microbial fermentation. Instead of the consistent and expensive minerals supplement required for Hfx. mediterranei DSM1411 growth, microfiltered seawater was added to the wasted bread-derived substrate. The suitable ratio of wasted bread homogenate and seawater, corresponding to 40:60, was selected. The addition of proteases and amylase to the bread homogenate promoted the microbial growth but it did not correspond to the increase of bioplastic production by the microorganism, that reach, under the experimental conditions, 1.53 g/L. An extraction procedure of the PHBV from cells, based on repeated washing with water, followed or not by a purification through ethanol precipitation, was applied instead of the conventional extraction with chloroform. Yield of PHBV obtained using the different extraction methods were 21.6 ± 3.6 (standard extraction/purification procedure with CHCl3:H2O mixture), 24.8 ± 3.0 (water-based extraction), and 19.8 ± 3.3 mg PHAs/g of wasted bread (water-based extraction followed by ethanol purification). Slightly higher hydroxyvalerate content (12.95 vs 10.78%, w/w) was found in PHBV obtained through the water-based extraction compared to the conventional one, moreover, the former was characterized by purity of 100% (w/w). Results demonstrated the suitability of wasted bread, supplemented with seawater, to be used as substrate for bioplastic production through fermentation. Results moreover demonstrated that a solvent-free extraction, exclusively based on osmotic shock, could be used to recover the bioplastic from cells.

Manipulation of Nitric Oxide Levels via a Modified Hydroxyethyl Starch Molecule

IntroductionAlthough the improving effect of nitric oxide (NO) donors has experimentally been demonstrated in shock, there are still no NO donor medications clinically available. Thiol-nitrosothiol-hydroxyethyl starch (S-NO-HES) is a novel molecule consisting of NO coupled to a thiolated derivative of hydroxyethyl starch (HES). It was aimed to assess the ability of S-NO-HES to serve as an NO donor under a variety of in vitro simulated physiologic conditions, which might be the first step to qualify this molecule as a novel type of NO donor-fluid. MethodsWe studied the effect of temperature on NO-releasing properties of S-NO-HES in blood, at 34°C, 37°C, and 41°C. Ascorbic acid (Asc) and amylase were also tested in a medium environment. In addition, we evaluated the activity of S-NO-HES in the isolated aortic ring and Langendorff-perfused heart setup. ResultsThe NO release property of S-NO-HES was found at any temperature. Asc led to a significant increase in the production of NO compared to S-NO-HES incubation (P < 0.05). The addition of amylase together with Asc to the medium further increased the release of NO (P < 0.05). S-NO-HES exerted significant vasodilatory effects on phenylephrine precontracted aortic rings that were dose-dependent (P < 0.01). Furthermore, S-NO-HES significantly increased the heart rate and additionally reduced the duration of the cardiac action potential, as indicated by a reduction of QTc-B values (P < 0.01). ConclusionsWe demonstrated for the first time that the S-NO-HES molecule exhibited its NO-releasing effects. The effectiveness of this new NO donor to substitute NO deficiency under septic conditions or in other indications needs to be studied.

Research Note: Amylase supplementation improves starch and amino acids digestibility of faba bean for broilers

This study aimed to investigate the effect of amylase on the nutritional value and nitrogen-corrected apparent metabolizable energy (AMEN) of faba bean (FB) seeds in broiler chicken nutrition. The study was conducted on a total of 84, one-day-old male broiler chickens (Ross 308). The birds were fed a reference corn-soybean meal diet form d 1 to 16, and they were divided into three groups (1st day). The first group (1) was fed the same reference diet as in the first part, whereas 50% of the reference diet was replaced by FB seeds in experimental groups (2 and 3). Group 3 was supplemented with amylase, which contained 0.14 g/kg dry matter (DM). At the end of the study period, excreta and digesta samples were collected from all groups to estimate the nutritional value. Some fraction of starch found in the seeds was part of starch which is resistant to digestion (143.9 g/kg DM). The addition of exogenous amylase to the diet increased the apparent ileal digestibility of DM, CP, and starch in FB seeds. The AMEN value increased as a result of enzyme addition (P < 0.05). The group 3 showed higher apparent ileal digestibility of essential amino acids (Lys, Leu, Phe, His) as well as nonessential amino acids (Tyr, Ala, Pro) in FB seeds. It could be concluded that FB seeds supplemented with exogenous amylase have a greater nutrition value for broiler chickens.

Corn kernel hardness, drying temperature and amylase supplementation affect live performance and nutrient utilization of broilers

Drying temperature (DT) of corn can influence its nutritional quality, but whether this is influenced by endosperm hardness is not clear. Two parallel experiments were conducted to investigate the effects of 2 yellow dent corn hybrids with average and hard kernel hardness, dried at 3 temperatures (35, 80, and 120°C), and 2 supplementation levels of an exogenous amylase (0, 133 g/ton of feed) on live performance, starch and protein digestibility, and energy utilization of Ross 708 male broilers. Twelve dietary treatments consisting of a 2 × 3 × 2 factorial arrangement were evaluated using 3-way ANOVA in a randomized complete block design. In Experiment 1, a total of 1,920 male-chicks were randomly allocated to 96 floor pens, whereas 480 day-old chicks were distributed among 96 cages for Experiment 2. At 40 d, interaction effects (P < 0.05) were detected on BWG, FCR, and flock uniformity. Supplementation with exogenous amylase resulted in heavier broilers, better FCR and flock uniformity, only in the diets based on corn dried at 35°C. Additionally, interaction effects were observed on FCR due to kernel hardness and DT (P < 0.01), kernel hardness and amylase supplementation (P < 0.001), and DT and amylase supplementation (P < 0.05). Exogenous amylase addition to the diets based on corn with an average hardness improved FCR up to 2 points (1.49 vs. 1.51 g:g) whereas there was no effect of amylase on FCR of broilers fed diets based on corn with hard endosperm. Total tract retention of starch was increased (P < 0.05) in broilers fed diets based on corn with average kernel hardness compared to hard kernel. Corn dried at 80 and 120°C had up to 1.21% points less starch total tract retention than the one dried at 35°C. Supplementing alpha-amylase resulted in beneficial effects for broiler live performance, energy utilization, and starch total tract digestibility results. Treatment effects on starch characteristics were explored. Corn endosperm hardness, DT and exogenous amylase can influence the live performance of broilers. However, these factors are not independent and so must be manipulated strategically to improve broiler performance.

Influence of Amylase Addition on Bread Quality and Bread Staling

In this study, the effect of two amylases (maltogenic amylase and maltotetraose-producing amylase) on the rheological properties of wheat flour as well as on the quality and staling of partially baked bread is investigated. The rheological measurements obtained from a Viscograph showed that both amylases reduced the peak viscosity, final viscosity, and setback viscosity of wheat flour, whereas maltotetraose-producing amylase significantly improved the specific volume of fresh bread. Furthermore, the addition of both types of amylases was able to reduce crumb firmness and amylopectin retrogradation, due to their ability to partially hydrolyze starch molecules and generate low molecular weight dextrin, which was confirmed by the analysis of the maltooligosaccharide composition of the breads. The results indicated that the effects of maltotetraose-producing amylase were more pronounced than those of maltogenic amylase in terms of slowing bread staling.

A physicochemical assessment of the thermal stability of dextrin\u2013colistin conjugates

Attachment of polysaccharide carriers is increasingly being used to achieve precision delivery and improved effectiveness of protein and peptide drugs. Although it is clear that their clinical effectiveness relies on the purity and integrity of the conjugate in storage, as well as following administration, instability of polysaccharide-based conjugates can reduce the protective efficacy of the polymer, which may adversely affect the bioactive’s potency. As a model, these studies used dextrin–colistin conjugates, with varying degrees of polymer modification (1, 2.5 and 7.5 mol% succinoylation) to assess the effect of storage temperature (− 20, 4, 21 and 37 °C) and duration (up to 12 months) on saccharide and colistin release and antimicrobial activity. Estimation of the proportion of saccharide release (by comparison of area under the curve from size exclusion chromatograms) was more pronounced at higher temperatures (up to 3 and 35% at − 20 °C and 37 °C, respectively after 12 months), however, repeated freeze–thaw did not produce any measurable release of saccharides, while addition of amylase (20, 100, 500 IU/L) caused rapid release of saccharides (> 70% total within 24 h). At all temperatures, conjugates containing the lowest degree of succinoylation released the highest proportion of free colistin, which increased with storage temperature, however no trend in saccharide release was observed. Despite the clear physical effects of prolonged storage, antimicrobial activity of all samples was only altered after storage at 37 °C for 12 months (> threefold decreased activity). These results demonstrate significant release of saccharides from dextrin–colistin conjugates during prolonged storage in buffered solution, especially at elevated temperature, which, in most cases, did not affect antimicrobial activity. These findings provide vital information about the structure–activity relationship of dextrin–colistin conjugates, prior to full-scale commercial development, which can subsequently be applied to other polysaccharide-protein and -peptide conjugates.

Influence of Enzyme Supplementation in the Diets of Broiler Chickens Formulated with Different Corn Hybrids Dried at Various Temperatures.

Simple SummaryThe use of exogenous enzymes is a common nutrition strategy of the poultry industry. However, the influence of this additive on the microbiota and its efficiency when the diets are formulated with different hybrids of corn dried under high temperature are still unclear. From a practical point of view, evaluating the mode of action of enzymes in different situations is crucial to ensure competitive performance results with low production costs. The current study confirmed that regardless of corn hybrids and drying temperature, dietary supplementation with amylase, xylanase, and protease was beneficial for intestinal morphology and allowed a modulation of the cecal microbiota. This influence may have changed the digestive process and use of nutrients by the broilers, resulting in better animal performance.We evaluated the influence of enzymatic supplementation on the growth performance and cecal microbiota of broilers. A total of 2160 1-day-old male chicks were used in a 3 × 2 × 2 factorial arrangement (three corn hybrids, two drying temperatures −80 and 110 °C, with or without the inclusion of an enzymatic blend (amylase, xylanase, and protease) (20 birds/pen, n = 9). For all performance and digestibility parameters, we observed, in general, isolated effects of the corn hybrids and drying temperature. Birds that received the enzymatic blend in the diet showed better weight gain from 1 to 21 days (d) and better digestibility coefficients of nutrients at 42 d. Birds fed diets with corn dried at 80 °C showed a better feed conversion ratio from 1 to 42 d. At 21 d of age, enzymatic supplementation had positive effects on jejunum morphology. Enzyme supplementation increased the abundance of the phylum Tenericutes, class Bacilli and Mollicutes, reduced Clostridia, and increased the abundances of the families Lactobacillaceae, Anaeroplasmataceae, and O_RF39;F. In conclusion, the addition of amylase, xylanase, and protease led to a better nutrient digestibility, performance, and intestinal morphology. In addition, enzyme supplementation changed the diversity, composition, and predicted function of the cecal microbiota at d 21.

The effect of amylase and cellulase enzymes on biogas production from rice husk waste using solid-state anaerobic digestion (SS-AD) method

Biogas comes from an anaerobic biological decomposition process. Biogas is an alternative energy that potential to be developed in Indonesia. One of the various raw materials that can be used in biogas production is rice husk waste. This study analyzes the effect of pretreatment using amylase and cellulase enzymes on the production of biogas from rice husk waste in the Solid-State Anaerobic Digestion (SS-AD) method. Rice husk waste was used as a substrate with a C/N ratio of 25%, and the total solids used was 21%. Rice husk is biomass that is difficult to degrade because it has high lignin content; therefore, pretreatment is necessary. One of them is biological pretreatment using enzymes. The enzymes used are amylase and cellulase enzymes with various concentrations of 9%, 12%, 15%, and 18%. The biogas produced was then measured every two days for 40 days of the study with the volume of biogas as the observed parameter. The results showed that pretreatment with the addition of amylase and cellulase enzymes could increase biogas production. The highest biogas production was obtained by pretreatment of 18% amylase enzyme, namely 1466 mL, and by pretreatment of 18% cellulase enzyme, namely 1075 mL.

Banana-based Biofuels for Combating Climate Change: How the Composition of Enzyme Catalyzed Solutions Affects Biofuel Yield

This investigation was inspired by the ever so precarious nature of climate change and its detrimental implications on our planet and society. One alternative form of energy that shows promise amongst others in alleviating the climate crisis, is biofuel. An extensive experiment was conducted to investigate whether amylase or yeast had a more prominent role in determining the bioethanol concentration and bioethanol yield of banana samples. Given that amylase facilitates the breakdown of complex polysaccharides into simple sugars, we hypothesized that amylase would have the most significant impact on the bioethanol yield and concentration of the samples. This was determined to be the case when the fermentation batch samples received enzyme compositions of yeast and amylase independently. However, when added concurrently, the optimal enzyme composition for maximizing the bioethanol concentration and yield was the addition of amylase and yeast in a 3:2 ratio. Thus, while yeast is an essential component for producing bioethanol, the proportion of amylase supplied through a joint amylase-yeast mixture has a more significant impact on the bioethanol yield. This provides some insight into what industrial engineers would need to take into consideration when mass-producing bioethanol on the industrial scale, namely that of the optimal enzyme composition with respect to cost and the bioethanol yield. Moreover, this study provides a greater understanding of the mechanisms and implications involved in enzyme-based biofuel production, specifically of those pertaining to amylase and yeast.

Formulation and processing of gruels made from local ingredients, thin enough to flow by gravity in enteral tube feeding

Designing enteral foods from local ingredients for tube feeding of low-income people who cannot eat orally is needed. Two processing methods, involving the addition of amylase or malt, were used to thin a blenderized tube feeding formula based on sorghum, sesame and soybean seeds. Two composite flours, either with higher carbohydrate (F1D) or higher lipid (F2D) contents were formulated to obtain an enteral food aimed at adults. To thin the formula enough for it to flow inside the feeding tube, increasing concentrations of amylase (0.27-2.17g/100g DM) were added to gruels F1D (F1DE) and F2D (F2DE) prepared at 25% DM. Sorghum malt was also added to F1D (F1DM) as an alternative source of amylase. But F1DE and F1DM flow times in a 50cm feeding tube (10 Fr) remained much longer (up to 14s) than that of the commercial enteral food (4s). The F1DE and F1DM osmolalities (485 and 599 mOsmol/Kg water, respectively) were higher compared to that of F1D but remained within the range specified for adult enteral food. F1D, F1DE and F1DM gruels showed pseudoplastic behavior. Their loss ratio (tan ), elastic (G') and loss (G'') moduli were similar, but apparent viscosity, flow time in the feeding tube and consistency index (k) showed that F1DE was thinner than F1DM. Adding an incubation step before cooking of F1DM suspension allowed further thinning of the gruel, showing it is possible to formulate an enteral food using local ingredients that flows by gravity in the feeding tube.

Enhancing nutrient utilization of broiler chickens through supplemental enzymes

The objective was to determine effects of adding phytase, amylase, and a cocktail of non-starch polysaccharide-degrading enzymes (NSPase) individually or in combinations to corn-soybean meal-based diet for broilers on apparent ileal digestibility (AID) of nutrients and dietary AMEn value. Four hundred and eighty male broiler chicks were divided into 80 groups and fed 8 diets in a completely randomized design (10 groups/diet) from day 15 to 21 of age. The diets were basal diet unsupplemented or supplemented with phytase (1,500 FTU/kg; Ronozyme HiPhos), amylase (80 KNU/kg, Ronozyme HiStarch), and NSPase (75g/metric ton, Ronozyme Multigrain) individually or in all possible combinations. The basal diet contained the phytase at 1,000 FTU/kg, and was formulated to meet the NRC (1994) recommended nutrient requirements for broiler chickens except for ME, Ca, and non-phytate P, which were reduced by 150kcal/kg, 0.18%, and 0.15%, respectively. Addition of phytase to the basal diet increased (P < 0.05) AID of P from 40.4 to 59.3%. Addition of amylase, NSPase or a combination of amylase and NSPase to the phytase-supplemented basal diet further increased (P < 0.05) AID of P to 63.4, 69.9, and 67.3%, respectively. Addition of phytase, amylase or a combination of amylase and NSPase to the basal diet did not affect dietary AMEn value. However, addition of NSPase alone or a combination of phytase and amylase or of phytase and NSPase to the basal diet improved (P < 0.05) dietary AMEn value from 3,203 to 3,339, 3,309 or 3,289kcal/kg, respectively. In conclusion, it is more beneficial (with regard to AID of P and dietary AMEn) to add amylase and NSPase to phytase-supplemented diets for broilers. Because the basal diet contained phytase at 1,000 FTU/kg, the increase in AID of P due to supplemental phytase (1,500 FTU/kg) indicates that supplemental phytase at 2,500 FTU/kg is more beneficial with regard to improving AID of P than supplemental phytase at 1,000 FTU/kg.

PH- and Amylase-Responsive Carboxymethyl Starch/Poly(2-isobutyl-acrylic acid) Hybrid Microgels as Effective Enteric Carriers for Oral Insulin Delivery.

Oral delivery of insulin has the potential to revolutionize diabetes care since it is regarded as a noninvasive therapeutic approach without the side effects caused by frequent subcutaneous injection. However, the insulin delivery efficiency through oral route was still limited, likely due to the chemical, enzymatic and absorption barriers. In this study, a novel type of pH- and amylase-responsive microgels as an insulin drug carrier for oral administration was developed to improve the drug delivery efficiency. The microgels were prepared via aqueous dispersion copolymerization of acrylate- grafted-carboxymethyl starch (CMS- g-AA) and 2-isobutyl-acrylic acid ( iBAA). The resulting hybrid microgels with the P iBAA contents of 13.6-45.3 wt% exhibited sharp pH-sensitivity, which was revealed by the changes in particle size of the microgels and the transmittance of the microgel aqueous solution. The accelerated decomposition of the CMS-containing microgels in response to amylase was demonstrated by chromogenic reaction and morphology change. Insulin was loaded into the microgels by swelling-diffusion method, and the insulin release from the insulin-loaded microgels in vitro was found to be triggered by pH change and addition of amylase, which was highly dependent on the microgel component. Cytotoxicity assay was performed to show the good biocompatibility of the microgels. In addition, the tests of cellular uptake by Caco-2 cells and transmembrane transport through the Caco-2 cell monolayers were carried out to confirm the intestinal absorption ability of the insulin-loaded microgels. Finally, the oral administration of insulin-loaded microgels to STZ-induced diabetic rats led to a continuous decline in the fasting blood glucose level within 2 to 4 h, and the hypoglycemic effect maintained over 6 h in vivo. The relative pharmacological availability of the insulin-loaded microgels was enhanced 23-38 times compared to free-form insulin solution through oral route. Therefore, the novel starch-based microgels may have potential as an efficient platform for oral insulin delivery.

Postruminal digestion of starch infused into the abomasum of heifers with or without exogenous amylase administration.

The effect of an exogenous amylase on postruminal digestion of starch infused into the abomasum of cattle was studied. Four rumen-cannulated heifers were fed 5.5 kg DM/d of a diet without starch, and assigned randomly to a crossover design. The experiment consisted of 2 periods lasting 23 d each with 10 d for adaptation to the diet followed by 13 d of abomasal infusion and sample collection. During the first 3 d of each infusion phase, isotonic saline solution was infused (1 liter/h) for measurement of baseline values in feces, followed by daily infusions of 880 g DM corn starch (1 kg/10 liters of water) without or with the addition of 2% of amylase. Titanium dioxide (10 g/d) was ruminally administered for estimation of fecal excretion. Digestion of starch in small intestine was calculated as the difference between the amounts of infused starch, disappeared from hindgut and fecal excretion. The apparent disappearance of starch from the hindgut was estimated based on the increment of microbial nitrogen (N) excretion due to starch infusion (1 g microbial N/100 g fermented starch) compared to baseline values. The concentration of purine bases in feces was used to estimate excretion of microbial N. Microbial N excretion increased with starch infusion (P < 0.05) but was not influenced by amylase (P = 0.81). Starch disappearance from the small intestine was not improved by amylase (P = 0.78) and averaged 85%. Amylase affected neither blood concentration of glucose (P = 0.80) nor of insulin (P = 0.26), but glucagon was lower without (P < 0.0001) than with amylase. The infusion of starch increased fecal excretion of total VFA (acetate, propionate, and butyrate) by 53% (P < 0.05), which indicates increased carbohydrate fermentation in the hindgut and incomplete digestion of starch in the small intestine. However, the excretion of total VFA was not affected by amylase (P = 0.66). Lactate excretion was higher at the second day of starch infusion (P < 0.05) without than with amylase, which suggests lower flow of starch from the small intestine to the hindgut due to a possible effect of amylase addition in animals not adapted to starch digestion. However, lactate excretion returned near to baseline values within 2 d, which was probably due to increase of lactate-utilizing bacteria and the adaptation of the microbial population in the hindgut. Further studies with higher starch levels and addition of amylase are recommended.