Enzyme Inclusion Research Articles

Diet supplementation with phytase and xylanase on laying performance, egg quality, apparent metabolizable energy, and phosphorous use in laying hens

Background and Aim: The inclusion of supplementary enzymes, such as phytase and xylanase, in poultry diets enhances nutrient digestibility and reduces excreta output, minimizing environmental impact. This study aimed to evaluate the effects of phytase and xylanase supplementation on laying performance, egg quality, apparent metabolizable energy (AME), and phosphorus utilization in laying hens. Materials and Methods: A total of 576 laying hens, aged 47 weeks, were randomly assigned to four dietary treatments: (T1) a control diet, (T2) a negative control diet with reduced energy and phosphorus, (T3) T2 supplemented with phytase (50 g/ton) and xylanase (100 g/ton), and (T4) T2 supplemented with phytase and NSP enzyme mix. The study was conducted over 12 weeks. Data on body weight, laying performance, egg quality, AME, and phosphorus utilization were analyzed using a one-way analysis of variance, with statistical significance set at p < 0.05. Results: No significant differences were observed in hen-day egg production, egg weight, or feed intake across groups. However, enzyme supplementation (T3 and T4) significantly reduced feed costs per kg egg and feed cost per egg compared to the control (p < 0.05). Eggshell strength improved significantly in T3 and T4 groups, while yolk color declined in these groups compared to T1 (p < 0.05). Other quality metrics, such as albumen height and Haugh units, showed no significant differences. Apparent metabolizable energy, phosphorus retention time, and feces scores were not significantly affected by dietary treatments. Conclusion: Phytase and xylanase supplementation in diets with reduced nutrient content effectively reduced production costs while maintaining laying performance and improving eggshell strength. These findings support the use of these enzymes as cost-efficient tools in poultry production without compromising egg quality. Keywords: egg quality, feed efficiency, laying hen, nutrient digestability, phosphorus utilization, phytase, poultry enzymes, xylanase.

Effect of two different dietary endo-1,4-β-xylanases on growth performance, intestinal histomorphology, caecal microbial population and short-chain fatty acid composition of broiler chickens

Effects of two different dietary xylanases on broiler performance and gut integrity were determined. Totally, 7600-d-old Ross-308 chicks were used for 41 d. Chicks were randomly assigned to 4 dietary treatments with 10 replicates (95 males, 95 females per replicate); they were: positive control (PC) based on Ross 308 nutrient specification; negative control (NC) with less ME (80 kcal/kg), crude protein and amino acids compared to PC; NC + AO with addition of xylanase derived from Aspergillus oryzae (AO); NC + TB with addition of xylanase derived from Trichoderma bisset (TB). Birds fed on PC diet significantly (p < 0.05) had higher final body weight (BW) and body weight gain (BWG) compared to other treatments. The final BWG and feed conversion ratio (FCR) were not fully compensated with the addition of the enzymes compared to PC, and FCR was found significantly (p < 0.05) higher in birds fed on diets NC + AO and NC + TB. However, dietary xylanase significantly improved FCR, villus surface area and total goblet cell number in NC + AO and NC + TB birds compared to NC. Xylanase supplementation significantly increased the concentrations of branched-chain fatty acid (BCFA) and total caecal short-chain fatty acid (SCFA) (p < 0.05). Furthermore, both xylanases increased lactobacillus count (p < 0.05). Dietary treatments had no effect on carcase parameters and mortality rate. In conclusion, along with improving performance parameters, the inclusion of xylanase enzymes with nutrient matrix value in broilers’ diet may ameliorate intestinal morphology, stimulate caecal microflora and increase caecal SCFA concentrations, but nutrient contribution of both enzymes seems overestimated. HIGHLIGHTS Dietary xylanase could not compensate the final BWG and FCR compared to positive control. Dietary xylanase increased villus surface area and total goblet cell number. Dietary xylanase increased the concentrations of branched-chain fatty acids and total caecal short-chain fatty acid.

The Biarylitides: Understanding the Structure and Biosynthesis of a Fascinating Class of Cytochrome P450 Modified RiPP Natural Products.

The biarylitides are a recently discovered class of RiPP natural products that are fascinating both from the small size of the core peptides as well as the diversity of peptide crosslinking exhibited by the cytochrome P450 enzymes found in these systems. In this review, we address the discovery and biosynthetic diversity of these systems and discuss the methods and challenges of analysing the structures of these constrained cyclic peptides. We also discuss the structures of the P450 enzymes involved in these pathways and address the potential for alternate catalytic outcomes and activities as seen most recently with the inclusion of biarylitide related enzymes within rufomycin biosynthesis.

CHANGES IN THE ELEMENTAL COMPOSITION OF THE CHYME AND METABOLIC PROCESSES IN THE RUMEN WHEN EXOGENOUS ENZYMES ARE USED IN FEEDING BULLS

Introduction. Coordination of the processes of scar digestion and metabolism in ruminants is a prerequisite for achieving high efficiency in the use of feed nutrients, improving product quality and profitability of production. The purpose of this study was to evaluate whether the inclusion of the exogenous lipase enzyme in the diet of bullocks in feedlots has a positive ef-fect on physiology, fermentation in the rumen, digestibility and the profile of fatty acids in the rumen. Material and methods. In the experiment, 8 animals were used, divided into 2 groups. the bulls of the control group received the basic diet (RR), the experimental group also received RR with the inclusion of the lipase enzyme at a dosage of 25 g / head / day. The level of volatile fatty acids (VFA) in the contents of the scar was determined by gas chromatography on a gas chromatograph "Crystallux-4000M", determination of nitrogen forms accord-ing to GOST 26180-84. The elemental composition of biosubstrates was studied using atomic emission and mass spectrometry (NPP-ISP and MS-ISP) in the IC of the Central Research Center of the FNC BST RAS. Results. The study of the ratio of LVH in the scar fluid showed that in the experimental group of animals the content of acetic acids decreased by 18.35%, propionic acid – 15.29%, butyric acid – 34.28%, valerian acid – 11.11%, while caproic acid was not found in the experimental group. Nitrogen metabolism in the body showed that the level of total nitrogen in the scar fluid increased by 64.71%, non–protein 53.66%, protein 66.22%, ammonia 78.57% (p 0.05), respectively. Analysis of the biomass of bacteria and protozoa showed that during the experiment, these indicators were higher in the experimental group of animals by 17.07% and 22.22%, respectively, compared with the control. The trace element composition of the scar fluid revealed differences in the mechanism of action of the lipase enzyme on the metabolism of chemicals. There was an increase in the concentration of chemical elements in the experimental group of bulls Ca – by 11.51%, P –17.72%, K – 20.34% (p≤0.05), Fe – 21.31%, Zn – 10.14%, Cu – 23.53%, Se – 39.13%. Against the background of a decrease in Ni – 11.44%, Ba – 14.67%, B – 9.00%, I – 33.33%, Al – 15.71 and Sr – 15.85% (p 0.05). Conclusion. Thus, the study of microbial communities of the LJ rumen, primarily of cattle, is very relevant due to the possibility of rapid diagnosis and prevention of many pathologies of the digestive system associated with improper feeding, which, as a rule, is accompanied by the development of pathogenic and opportunistic microorganisms.

PSV-27 Optimizing corn silage utilization in beef feedlot diets: Evaluating the impact of exogenous glucoamylase addition on performance and digestibility

Abstract The addition of an exogenous glucoamylase enzyme preparation to feedlot diets that contain a high percentage of corn silage may improve digestibility and yield positive outcomes in growth and efficiency. However, it is unclear what concentration of corn silage in the diet would most benefit from exogenous glucoamylase. Crossbred steers [n = 180; body weight (BW) = 459 ± 11.8 kg] were divided into 30 pens (5 pens per treatment; 6 steers per pen) and assigned to one of six treatments arranged as a 3 x 2 factorial investigating 3 forage concentrations (15, 42, and 69% corn silage) and 2 exogenous glucoamylase additions (0 or 183 enzyme units/kg of dry matter per d). Steers were fed 42% corn silage, 30% corn, 23% dried distillers grains with solubles, and 5% supplement at feedlot entry and for 2 wk prior to initiation of the study. Steers on the 15% and 69% corn silage diets were transitioned to treatment diets by wk 3 of the study (d 15), with the first transition occurring on d 0. Diet transitions occurred once weekly. The animals were on feed for 105 ± 10.4 d and had ad libitum access to feed and water. Average daily gain (ADG), dry matter intake (DMI), and gain:feed were assessed both overall and for two distinct periods: d 0 to 50, and d 51 until slaughter. Data analysis was performed using the GLIMMIX procedure of SAS. From d 0 to 50, ADG, DMI, and gain:feed did not differ due to forage concentration or enzyme inclusion and there was no interaction (P ≥ 0.39). For d 51 to slaughter, enzyme inclusion interacted with forage concentration such that the enzyme improved ADG (P = 0.0003) and gain:feed (P = 0.01) for steers fed the 42% corn silage diet but did not effect ADG and gain:feed for steers fed the 15 or 69% corn silage diets. For d 51 to slaughter DMI was not affected by forage concentration, enzyme inclusion, or their interaction (P ≥ 0.21). When calculated for the entire study (d 0 to slaughter), increasing forage concentration decreased ADG and gain:feed (P ≤ 0.0006), but did not effect DMI (P = 0.22). Overall, enzyme addition did not impact ADG, DMI, or gain:feed and did not interact with forage concentration (P ≥ 0.50). Carcass characteristics were not affected by forage concentration, enzyme addition, or their interaction (P ≥ 0.19). Overall results suggest that an exogenous glucoamylase enzyme improves growth and efficiency of heavier feedlot cattle fed moderate amounts of forage later in the feeding period.

497 Amylase supplementation influences nutrient digestibility regardless of corn type in broiler chicken diets

Abstract Corn is the primary energy supply in the broiler chicken diet. Physical and nutritional characteristics related to corn endosperm can affect nutrient utilization, as well as the inclusion of exogenous enzymes. This study aimed to evaluate two different corn types classified according to endosperm traits in diets for broiler chickens supplemented or not with amylase. Male broiler chickens (n = 360; 1-d old Cobb 500) were allocated in metabolic battery cages and randomly assigned in a 2 x 2 factorial arrangement (2 corn types; 2 amylase doses: 0 and 90 g/t of diet). A hardness test was performed to categorize and classify each corn type according to breakage force. Corn endosperms were assessed through scanning electron microscopy to investigate structure variations of starch granules. From 19 to 21 d, excreta samples were collected to estimate apparent total tract retention (ATTR) of nutrients and apparent metabolizable energy (AME). On d 26, ileal digesta content was collected to determine apparent ileal nutrients digestibility (AID) and ileal digestible energy (IDE). Data were analyzed using the Student t-test for hardness test (P &amp;lt; 0.05). The Tukey test was performed with a significance declared of 5% to compare the averages obtained for ATTR and AID using the GLM procedure on R Software. Differences were observed in the hardness test (P &amp;lt; 0.05), which classified each corn type either as hard (217 N) or soft (166 N; Table 1). The SEM highlighted structural variations between the floury and vitreous endosperm of each corn. In soft corn, starch granules were more dispersed in the floury endosperm, with a less dense distribution observed in the vitreous endosperm compared with hard corn (Figure 1). However, hard corn exhibited a more compact starch granules distribution in both floury and vitreous endosperm. No differences were observed (P &amp;gt; 0.05) in ATTR of dry matter, crude protein, or AME attributed to corn type, but amylase supplementation improved ATTR of dry matter (P = 0.015) and crude protein (P = 0.020; Table 2). On AID, corn type affected (P = 0.041) dry matter coefficient, and amylase supplementation significantly improved dry matter (P = 0.019), crude protein ileal digestibility (P = 0.012), and IDE (P = 0.018). No interactions between corn type and amylase supplementation were observed (P &amp;gt; 0.05). These findings demonstrated that amylase supplemented in broiler diets impacts nutrient utilization regardless of the corn type used.

GROWTH PERFORMANCE OF BROILER CHICKENS FED DIET SUPPLEMENTED WITH BIONIC PREBIOTIC (HILYSES) AND HARDCORE YEAST CELL WALL

In a bid to boost productivity, several dietary manipulations have been carried out on poultry, ranging from inclusion of growth promoters, hormones, organic acids, enzymes (Chuka and locally sourced additives Thus, this study was carried out to assess supplementation of regular diet of broiler chickens with bionic prebiotic (Hilyses) and hardcore Yeast Cell Wall (Immuno Wall). The experiment utilized a Completely Randomized Design (CRD) with 120 unsexed day-old chicks randomly allocated into four treatment groups (T1, T2, T3, and T4), each consisting of 30 chicks and replicated three times. The initial live weight of the broiler chicks at day old was recorded. The birds were weekly weighed to determine the live weight and weight gain. The quantity of feed offered to the birds daily and the left over the next morning were measured to ascertain feed intake. Mortality was noted. Over the 56-day experimental period, the results demonstrated notable improvements in various performance traits attributable to the inclusion of prebiotic and yeast supplements, except for the initial weight, which exhibited not statistically significant (P&gt;0.05) difference among the treatment groups. Significant (p&lt;0.05) differences were observed in the final weight, with birds fed diet 4 displaying the highest final weight, reaching 2680.85g. In contrast, the final weights of birds fed diet 3 and 2 were 2525.55g and 2450.10g, respectively, while those on diet 1 had the lower final weight of 2407.65g. It is worth noting that as the inclusion levels of prebiotic and yeast increased across the dietary treatments, a significant (p&lt;0.05) enhancement in final weight was consistently observed. The findings of this study unequivocally demonstrate the favourable growth performance of the treatment diets across all parameters examined. Specifically, it is evident that T4, with a 0.75% inclusion of prebiotics and yeast cells, yielded the highest final bod weight than both the other trial diets and the control diet. These findings highlight the potential for enhanced broiler chicken performance and overall health when incorporating these dietary additives.

Antioxidant Marine Hydrolysates Isolated from Tuna Mixed Byproducts: An Example of Fishery Side Streams Upcycling.

The aim of this research is to propose simple and scalable processes to obtain bioactive peptides extensively hydrolyzed starting from a tuna mixed biomass. The upcycling of this powdered biomass is challenging since it comes from the unsorted industrial side streams of the tuna canning process (cooked residues from fillet trimming) after a patented mild dehydration useful for preventing its degradation until its exploitation. Two different protocols were proposed, with and without the inclusion of an exogenous enzyme (Enzymatic-Assisted Extraction, EAE), with no relevant differences in yields (24% vs. 22%) and a comparable amino acid composition. Nevertheless, the former protocol (with EAE) provided peptides with an average molecular weight of 1.3 kDa, and the second one (without EAE) provided peptides with an average molecular weight of 2.2 kDa. The two corresponding types of tuna protein hydrolysates (Enzymatic Hydrolysates (EH) and Non-Enzymatic Hydrolysates (NEH)) were characterized by proximate compositions, pH, color profile, amino acid analysis, FTIR spectra, and molecular weight distribution. In addition, several biological analyses were performed to assess their potential use as nutraceutical supplements: special attention has been paid to antioxidant activity using three different methods to quantify it. EH showed the most promising antioxidant activity which could be exploited also in other fields (e.g., biomaterials, cosmetics).

Evaluation of black soldier fly larvae and protease supplementation of Lohmann Brown-lite laying hens performance and egg quality

Abstract Feed costs represent up to 60-70% of the total cost of poultry production, with protein sources being the most expensive feed component. Black soldier fly larvae meal (BSFLM) shows promise as a sustainable protein source for use in animal diets. This research was conducted to determine the effects of including up to 13% defatted BSFLM and a supplementary protease-based enzyme (Concentrase-P; CBS Bio Platforms, Inc., Calgary AB, Canada) on laying hen performance and egg quality. To test this objective, 180 Lohmann Brown-Lite hens (52 weeks of age) were fed experimental diets containing 0%, 6.5%, or 13% BSFLM, with or without Concentrase-P (3 × 2 factorial arrangement of treatments). The treatments were fed over 20 weeks. Results indicated reduced feed intake and body weight in laying hens fed a 13% dietary inclusion level of BSFLM (), while other performance traits (egg production and FCR) were similar among treatments () and met industry performance standards. Shell weight and egg breaking strength did not significantly change with the dietary inclusion level of BSFLM or enzyme inclusion (), although a significant decrease in egg weight and albumen height was noted (), as well as a significant decrease in albumen height for the Concentrase-P treatments (). Yolk colours were significantly lighter and redder in birds fed diets containing high levels of BSFLM (). Results support the use of BSFLM at dietary inclusion levels up to 13% in laying hen diets. Concentrase-P does not appear to be required in combination with BSFLM in terms of growth and laying performance, although potential benefits to other physiological factors are worth consideration.

Correlation between acute muscle damage and oxidative protection enzymes during different aerobic exercises

Different types of aerobic exercise can cause different disorders of homeostasis. This cross-over experiment aimed to determine the muscle fatigue and the antioxidative protection of female basketball players following a load caused by three different aerobic-type exercises (low-intensity continuous, high-intensity continuous, and high-intensity interval training). Twelve female basketball players (age 17.7±4.3 years; weight 67.3±9.8 kg; height 178.0±7.4 cm) voluntarily participated in the study. A wash-out period of 7 days between single sessions of different training was provided. Venous blood was drawn right before and immediately after each exercise session. The parameters that were analyzed are markers of muscle damage and enzymes of antioxidant protection. As a marker of muscle damage, myoglobin (F=2.884; p=0.065) and lactates (F=5.254; p=0.008) have higher values and statistically significant differences between training types. Creatinine shows higher values after each training session (F=4.053; p=0.022). Results of enzyme activity for oxidative protection show statistically significant differences between groups for catalase (F=5.811; p=0.005) with different types of training intervention. At the beginning of the preparatory period, parameters of acute muscle damage values are high. During the season, in response to different types of training, those parameters decrease in response to the body's adaptation to exercise-induced stress. Training leads to maintenance of physiological balance in the body and oxidative stress is not a necessary phenomenon of high aerobic training load. The inclusion of antioxidant protection enzymes decreases as the body adapts to a certain type of exercise.

113 Evaluation of high soybean meal diets with inclusion of dietary enzymes on the effect of growth performance and greenhouse gas emissions

Abstract Crossbred gilts [n = 48; initial body weight (BW) = 52.55 ± 4.12 kg] were used to evaluate the effects of high soybean meal (SBM) inclusion with dietary enzymes on the growth performance, nitrogen balance, and gas emissions of growing pigs. This study consisted of two replicates of 24 gilts placed in individual metabolism stalls. Pigs were randomly assigned to one of six dietary treatments and blocked by BW (4 pigs•treatment-1•replicate-1). Pigs were fed a common diet from d 0 to 12, the experimental diets were fed in two phased (d 12 to 24 and d 24 to 36). The experiment consisted of 3 collection periods, with each period subdivided into two subperiods to collect samples for gas emissions and nutrient balance. Manure samples were collected for 72 h. Twice daily, urine and feces were weighed, and a ratio of the excreta were subsampled to measure gas emissions. Measurement of nutrient digestibility and retention consisted of urine and feces collections over a 72 h-period. The six dietary treatments include: 1) basal control diet (NC) corn-soy diet supplemented with synthetic amino acids, 2) high soybean meal (HSBM) diet (PC) formulated to replace all synthetic lysine, 3) HSBM diet with xylanase (Ronozyme WX 5000; DM; WX), 4) HSBM diet with inclusion of a multi-enzyme additive including hemicellulase and pectinase (Ronozyme VP, DM; VP), 5) HSBM diet with inclusion of xylanase (Ronozyme WX 5000, DM) and multi-enzyme additive including hemicellulase and pectinase (Ronozyme VP, DM; WXVP), and 6) HSBM diet with 1,500 FYT/kg of phytase (HiPhorious, DM; SD). The NC, PC, WX, VP, and WXVP diets contained phytase (HiPhorious, DM Nutritional Products, Parsippany, NJ) to provide 600 phytase units (FTU) per kg of diet and was assumed to release 0.15% available phosphorous. The SD diet contained 1,500 FTU/kg and was assumed to release 0.21% available phosphorous. Additionally, high soybean meal diets within each phase were isocaloric. Data were analyzed as repeated measures in SAS 9.4 (Inst., Cary, NC) with fixed effects of treatment, collection period, and block. Pig was the experimental unit, and results were considered significant at P ≤ 0.05 and a tendency at 0.05 &amp;lt; P ≤ 0.10. Pigs fed VP had the greatest average daily gain (ADG) compared with pigs fed SD (P = 0.865; Table 1). Pigs fed VP had the greatest gain to feed ratio (GF) efficiency compared with pigs fed WXVP (P = 0.3544). Manure pH, urine pH and hydrogen sulfide tended to decrease in all treatments from phase 2 to phase 3. Pigs fed PC had the lowest manure pH compared with pigs fed SD (P = 0.0826; Table 2). In conclusion, the feeding of HSBM or adding select enzymes did not alter gas emissions or growth performance metrics.

Effect of commercial fibrolytic enzymes application to normal- and slightly lower energy diets on lactational performance, digestibility and plasma nutrients in high-producing dairy cows.

The inclusion of fibrolytic enzymes in the diet is believed to have positive effects on animal production. Hence, the objective of this study was to investigate the impact of supplementing diets with a commercial fibrolytic enzyme preparation (Vistamax; mixture of xylanase and cellulase) derived from Trichoderma reesei on lactational performance, digestibility, and plasma nutrient levels in high-producing dairy cows. Two dietary energy levels were considered: a normal energy diet (metabolizable energy = 2.68 Mcal/kg) and a slightly lower energy diet (metabolizable energy = 2.55 Mcal/kg). A total of 120 lactating Holstein cows (parity = 2; Days in Milk = 113 ± 23) were randomly assigned to four treatment groups using a 2 * 2 factorial arrangement. The dietary treatments consisted of: (1) normal energy diet without enzyme supplementation (NL); (2) normal energy diet with enzyme supplementation (NLE); (3) slightly lower energy diet without enzyme supplementation (SL); and (4) slightly lower energy diet with enzyme supplementation (SLE). The amount of enzyme added to the diets was determined based on previous in vitro studies and supplier recommendations. The enzyme and premix were mixed prior to the preparation of the total mixed ration, and the trial lasted for a duration of 42 days. The results indicated that the application of the fibrolytic enzyme did not have a significant effect on dry matter intake (DMI), but it did enhance the digestibility of dry matter (DM), neutral detergent fiber (NDF), potentially digestible NDF (pdNDF), organic matter (OM), milk production, milk urea nitrogen (MUN), and blood urea nitrogen (BUN). On the other hand, the slightly lower energy diet resulted in a decrease in DMI, milk production, milk protein yield, plasma free amino acids (FAA), and an increase in plasma B-hydroxybutyrate (BHBA). In conclusion, the inclusion of the fibrolytic enzyme in the diets of dairy cows led to improvements in the digestibility of DM, NDF, pdNDF, OM, milk production, and feed efficiency. Furthermore, the application of the enzyme to the slightly lower energy diet resulted in milk production levels comparable to those observed in cows fed the untreated normal energy diet.

Recombinase-Aided Loop-Mediated Isothermal Amplification on Human Plasmodium knowlesi.

Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique that can amplify specific nucleic acids at a constant temperature (63-65°C) within a short period (<1 hour). In this study, we report the utilization of recombinase-aided LAMP to specifically amplify the 18S sRNA of Plasmodium knowlesi. The method was built on a conventional LAMP assay by inclusion of an extra enzyme, namely recombinase, into the master mixture. With the addition of recombinase into the LAMP assay, the assay speed was executed within a time frame of less than 28 minutes at 65°C. We screened 55 P. knowlesi samples and 47 non-P. knowlesi samples. No cross-reactivity was observed for non-P. knowlesi samples, and the detection limit for recombinase-aided LAMP was one copy for P. knowlesi after LAMP amplification. It has been reported elsewhere that LAMP can be detected through fluorescent readout systems. Although such systems result in considerable limits of detection, the need for sophisticated equipment limits their use. Hence, we used here a colorimetric detection platform for the evaluation of the LAMP assay's performance. This malachite green-based recombinase-aided LAMP assay enabled visualization of results with the naked eye. Negative samples were observed by a change in color from green to colorless, whereas positive samples remained green. Our results demonstrate that the LAMP assay developed here is a convenient, sensitive, and useful diagnostic tool for the rapid detection of knowlesi malaria parasites. This method is suitable for implementation in remote healthcare settings, where centralized laboratory facilities, funds, and clinicians are in short supply.

Effect of addition of yeasts and enzymes during fermentation on physicochemical quality of fine aroma cocoa beans

Theobroma cacao is a recognized crop worldwide within the market for raw materials or processed products. Cocoa beans quality, industrial transformation and health benefits have been widely studied, but there are few studies on the fermentation process and even less on Ecuadorian cacao. In Latin America, this crop plays a very important role from an ecological and social viewpoint. Production is usually carried out by local farmers who also perform the natural fermentation of the beans, without specific management, which represents a challenge in terms of reliability and batch-to-batch homogeneity. This drawback can be effectively addressed by controlling the fermentation conditions. In addition, certain enzymes have the potential to improve the fermentation process. The aim of this work was to study the effect of using yeasts and/or enzymes during fermentation on the physicochemical quality characteristics of cocoa beans. Two trials were carried out, the first used yeasts (Saccharomyces cerevisiae), with and without molasses, and the second used enzymes (Polyphenoloxidase and Pectintranseliminase). During the fermentation process there was not external temperature control, although cocoa beans mass temperature was assessed. After the assay, the cutting test and physicochemical variables (percentage of testa, testa and cotyledon pH, moisture, ash, acidity, protein and fat contents) were evaluated. The yeasts trial stood out as the most favorable option, showing better fermentation in the cutting test, a slight increase in protein content and lower moisture. The inclusion of enzymes in the trials resulted in higher fermentation temperatures and fat content of the beans, but the samples showed lower fermentation percentage, higher moisture and mold presence, as enzymes make it difficult to completely dry the samples. It can be concluded that the addition of yeasts and/or enzymes influences fermentation and could improve the process in order to obtain high quality cocoa beans.

Effects of yeast and exogenous fibrolytic enzyme inclusion in the diet of hair lambs on performance, carcass traits, physicochemical parameters and meat fatty acid profile

Effects of yeast and exogenous fibrolytic enzyme inclusion in the diet of hair lambs on performance, carcass traits, physicochemical parameters and meat fatty acid profile

Nanoplate-based digital PCR for highly sensitive pork DNA detection targeting multi-copy nuclear and mitochondrial genes

The inclusion of ingredients derived from pigs in highly processed consumer products poses a significant challenge for DNA-targeted analytical enforcement, which could be overcome by using digital PCR. However, most species detection methods use digital PCR to target single-copy nuclear genes, which limits their sensitivity. In this work, we examined the performance of a nanoplate-based digital PCR method that targets multi-copy nuclear (MPRE42) and mitochondrial (Cytb) genes. Poor separation of positive and negative partitions, as well as a ‘rain effect’ were obtained in the porcine-specific MPRE42 assay. Among the optimization strategies examined, the inclusion of restriction enzymes slightly improved the separation of positive and negative partitions, but a more extensive ‘rain effect’ was observed. The high copy number of the MPRE42 amplicon is hypothesized to contribute to the saturation of the positive signal. In contrast, the porcine-specific Cytb assay achieved perfect separation of positive and negative partitions with no ‘rain effect’. This assay can detect as little as 0.4 pg of pork DNA, with a sensitivity of 0.05% (w/w) in a pork-chicken mixture, proving its applicability for detecting pork in meat and meat-based products. For the MPRE42 assay, potential applications in highly degraded products such as gelatin and lard are anticipated.

CREATION AND PROPERTIES OF BIOCOMPOSITE NANOPARTICLES BASED ON FUCOIDAN AS CARRIERS OF FIBRINOLYTIC ENZYME

Magnetically guided biodegradable nanocomposites are a new class of functional materials that expand the possibilities of clinical therapy. The paper describes the chemistry of the formation of a nanobiocomposite material based on a natural polymer synthesized by electrostatic interaction of a polysaccharide and magnetite in an aqueous medium. The possibility of electrostatic immobilization of a model fibrinolytic enzyme was studied. An increase in the electrochemical potential of particles during immobilization of a fibrinolytic enzyme was established by the method of dynamic light scattering. The degree of enzyme inclusion was 9.95±0.05 mass%. The average particle size of the nanobiocomposite material after immobilization of the enzyme was 264.3 nm. The study of the magnetic properties showed the absence of residual magnetization, which makes it possible to exclude the possibility of magnetic aggregation when using the biocomposite in a biological application. The saturation magnetization of the formed material was 57.1 ± 2.3 kA/m, which is lower than that of pure magnetite sol (MS ∼ 73.8 ± 3.4 kA/m), but sufficient for manipulation under conditions of resistance to blood flow. The kinetics of the release of an enzyme preparation in a phosphate buffer from a nanocomposite carrier based on fucoidan was studied. To study the kinetics of enzyme release by mathematical modeling, zero and first order models, the Korsmeier-Peppas model, the Higuchi model, and the Baker-Lonsdale model were used. It has been established that the kinetics of fibrinolytic enzyme release is described with high accuracy (r2 = 0.98) by the Korsmeier–Peppas equation, where the process of release of the enzyme preparation is influenced by diffusion obeying Fick's law. It has been established that zero and first order models are applicable only to the initial (0-40 min, where r2 = 0.96) and final (80-320 min, r2 = 0.93) stage of release.

Inclusion of exogenous enzymes in feedlot cattle diets: Impacts on physiology, rumen fermentation, digestibility and fatty acid profile in rumen and meat

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.

Impact of enzyme inclusion on the digestibility of protein, amino acids, calcium and phosphorus in growing pigs depending on meal size, frequency and sampling period

When digestibility trials are conducted, pigs are typically fed twice a day with a restricted amount of feed. These conditions are not representative of the feeding conditions in commercial farm. The present study aimed to determine the effects of meal size and frequency, xylanase and phytase inclusion as well as digesta sampling period, on the digestibility of protein, amino acids (AA), calcium (Ca) and phosphorus (P). Pigs (n = 6) were fitted with a T cannula, and each received all treatments following a 2 × 3 factorial arrangement in a 6 × 6 Latin square experimental design. The diets were supplemented or not with a combination of xylanase and phytase and divided into 3 feeding programs: one received 2 meals per day that met 3 times the maintenance energy requirement (2M), one received the same quantity of feed in 8 meals (8M), and another received an amount that supplied 5 times the maintenance energy requirements in 8 meals (8M+). Digesta was sampled from 0800 to 1200, 1200–1600, 1600–2000 and 0800–2000. The results indicated an improvement in the coefficient of apparent ileal digestibility of dry matter, crude protein, P, Ca, Val and Thr with enzyme inclusion in all meal distribution (enzyme effect, P < 0.05). Apparent ileal digestibility of Ca in the 0800–1200 period was reduced in 8M+ treatment compared to 2M and 8M whereas no difference was observed for meal size or frequency in 1200–1600 and 1600–2000 periods (Interaction meal × period, P = 0.017). A meal effect was observed for coefficient of apparent total tract digestibility of Ca (meal effect, P = 0.003) which was higher by 8% in pigs fed 8M compared to 8M+ (contrast 8M vs 8M+, P = 0.040). Saturation of the absorption processes may explain lower digestibility with 8M+ . Enzyme efficacy was not affected by meal size or frequency indicating minor modifications of digestive conditions. An 8-h sampling period would allow collection of a representative sample and avoid underestimating or overestimating digestibility of certain nutrients.

Optimizing techno-functionality of germinated whole wheat flour steamed bread via glucose oxidase (Gox) and pentosanase (Pn) enzyme innovation

Germination, a powerful biofortification technique, holds immense potential in bolstering the micronutrient profile of essential staple grains, thereby paving the way for optimal nutritional enhancement. The primary goal of this study was to improve the technological functionality of germinated wheat flour by incorporating pentosanase (Pn) and glucose oxidase (Gox) enzymes, with particular emphasis on the evolutionary changes in its components. The inclusion of Gox did not produce any substantial impact on the volumetric characteristics of the steamed bread. The incorporation of Pn and Gox has been seen to enhance the overall excellence of steamed bread by optimizing loaf volume and textural characteristics while also improving the thermal stability of the dough. The existence of two endothermic peaks could be attributed to bound water or alterations in the granules within the starch crystallization region. Adding Pn and Gox reduced and increased the formation and stability time of the dough, respectively. A certain ratio was employed to assess alternations in the crystallinity of starch granules over a limited range. After steaming, a significant decrease in IR1047/1022 was observed, indicating that the elevated temperature partially disrupted the internal starch crystal structure, leading to a gelatinization reaction with water. The ratio of tensile resistance (R) and elongation (E) of dough increased significantly compared to the control. The results obtained from this study indicate that the simultaneous inclusion of enzymes (Pn + Gox) holds significant promise for expanding the technological functionality of germinated wheat flour dough and improving the quality attributes of steamed bread.