Dietary Protease Research Articles

Effect of protease supplementation on amino acid digestibility of soybean meal fed to growing-finishing pigs in two different ages.

This study was conducted to investigate the effect of protease inclusion level in two different ages on the apparent (AID) and standardized (SID) ileal digestibility of crude protein (CP) and amino acids (AA) in soybean meal (SBM) fed to growing-finishing pigs. Ten cannulated pigs (21kg ± 2kg) were assigned to experimental diets in a duplicate 5 × 5 Latin square design. In Phase I (23 to 30 kg-pigs, 90 ± 17 days of age), ileal digesta was collected in 5 periods of 7d (5d adaptation and 2d ileal digesta collection). In phase II, (50 to 65 kg-pigs, 140 ± 17 days of age), ileal digesta was collected in 5 more periods of 7 d. For both phases, a corn starch-based diet was formulated with SBM as the sole source of CP and AA and containing titanium as an indigestible marker. Protease was supplemented at 0, 15,000, 30,000, and 45,000 NFP/kg of feed (0, 25, 50, and 75g/ton of ProAct 360®). A nitrogen-free diet was used to estimate basal ileal endogenous AA losses. Pigs were fed at 4% of their body weight, which was adjusted at the end of each period. Orthogonal polynomial contrasts were used to determine the linear and quadratic effects of dietary protease supplementation in each phase. In phase I, increasing levels of protease resulted in a linear increase (P<0.10) in SID for the 7/11 indispensable AA (Except Arg, His, Met + Cys, and Trp) and the average of all dispensable AA. In phase II, the SID of Ile, Leu, Met, Met + Cys, Val, the average of all indispensable AA, and 4/7 dispensable AA were quadratically increased (P<0.10). In most cases, supplementation with 30,000 NFP/ kg of feed (50g/ton) resulted in the greatest increase in AA digestibility. However, the linear response in phase I for some AA suggests that diets for younger pigs could be supplemented with a greater level (45,000 NFP/kg or 75g/Ton of feed). Interestingly, younger pigs had consistently increased (P<0.10) SID of CP and 15/18 AA (Except Arg, Cys, and Ser), being ~5.6% greater for indispensable AA when compared to older pigs. In conclusion, dietary protease supplementation can increase the SID of AA in soybean meal in both growing and finishing periods. Pig age can potentially influence AA digestibility, possibly related to a greater ileal endogenous AA flow in younger pigs. However, this fact warrants further investigation.

PSI-29 Enhancing growth performance and reducing environmental impact with dietary protease supplementation in growing pigs fed high- and low-density diets

Abstract Exogenous protease has emerged as a promising solution for enhancing protein digestion and utilization in animal diets, offering potential benefits for both animal performance and environmental impact mitigation in pig farming. In a 6-wk trial involving 140 growing pigs [initial body weight (BW) = 24.10 ± 1.66 kg], the effects of dietary protease supplementation on growth performance, nutrient digestibility, and noxious gas emission were investigated. Pigs were randomly assigned to one of four dietary treatments in a 2 x 2 factorial design with two levels of nutrient density and 0 or 125 g/ton supplementation of protease. The pigs were blocked by BW and housed in 28 pens with each pen consisting of 5 pigs (3 barrows and 2 gilts). The experimental design employed a randomized complete block design with 7 replicate pens per treatment. The data were analyzed using two-way ANOVA, with differences between means assessed using Tukey’s test (P &amp;lt; 0.05). Results revealed no significant interaction (P &amp;gt; 0.05) between nutrient density and protease supplementation concerning growth performance, nutrient digestibility, and noxious gas emission. Pigs subjected to high-density diets exhibited greater (P &amp;lt; 0.05) average daily gain (ADG) and gain-to-feed ratio (G:F) compared with those on low-density diets. While protease supplementation did not impact ADG and average daily feed intake (ADFI), there was a tendency towards reduced (P = 0.08) G:F throughout the 6-wk period. Pigs fed high-density diets demonstrated significantly greater (P &amp;lt; 0.05) apparent total tract digestibility (ATTD) of gross energy (GE) compared with their counterparts on low-density diets. Furthermore, the inclusion of protease significantly increased (P &amp;lt; 0.05) ATTD of GE. However, no discernible differences were noted among diets regarding the ATTD of dry matter and nitrogen. Regarding noxious gas emission, protease supplementation did not significantly affect total mercaptans and hydrogen sulfide (H2S) emission. Nonetheless, there was a notable trend towards reduced (P = 0.06) ammonia gas (NH3) emission with protease inclusion. Similarly, nutrient density did not exhibit a significant effect (P &amp;gt; 0.05) on noxious gas emission. In conclusion, the results of the study underscore the importance of nutrient density and protease supplementation in optimizing pig performance and environmental sustainability in swine production systems. Further research is warranted to elucidate the underlying mechanisms and refine strategies for maximizing the benefits of these dietary interventions.

375 The effects of protease supplementation on weaned piglet performance and intestinal function fed greater amounts of lower digestible protein sources

Abstract The present study investigated the impact of dietary protease supplementation on the growth performance and intestinal function of weaned piglets when fed a diet comprising high levels of low digestible protein sources, in comparison with a standard commercially representative piglet diet. A total of 180 weaned piglets, 21 d old, of Duroc × Large White × Landrace crossbreeds, with an initial body weight (BW) of 6.27 ± 0.45 kg, were randomly assigned to three dietary treatments: 1) a standard commercial positive control (PC) diet comprising 22.2% soy protein, 5% whey protein, and 3% fish meal, 2) a negative control (NC) diet comprising 30.1% soy protein without whey protein and fish meal, and 3) a NC diet supplemented with 0.02% protease (NC + PROT). The diets were formulated to be iso-nitrogenous and iso-caloric and were pelleted at conditions of 0.4 MPa and 75°C. The experimental design employed a completely randomized design with 6 replicate pens per treatment, each pen containing 10 piglets. Statistical analysis was conducted using one-way ANOVA, with differences between means assessed using Duncan’s Multiple Range Test (P &amp;lt; 0.05). In terms of piglet performance, feeding the NC diet, containing high levels of low digestible protein sources, resulted in a significant decrease (P &amp;lt; 0.05) in final BW, average daily gain (ADG), and average daily feed intake (ADFI) by 3.1%, 8.2%, and 5.2%, respectively, compared with piglets fed the PC diet. Conversely, the NC diet significantly increased (P &amp;lt; 0.05) the feed-to-gain ratio and incidence of diarrhea by 3.4% and 88.3%, respectively, relative to PC. However, the supplementation of protease in the NC + PROT diet restored final BW, feed-to-gain ratio, and incidence of diarrhea to the same level as PC, while ADG and ADFI were increased (P &amp;lt; 0.05) by 4.6% and 4.3%, respectively. Regarding intestinal function, relative to PC, piglets fed the NC diet exhibited significantly less (P &amp;lt; 0.05) duodenal villus height, crude protein digestibility, and gene expressions of duodenal peptide transporter PepT1 and jejunal amino acid transporter b0,+AT, while blood urea nitrogen and diamine oxidase concentrations were greater (P &amp;lt; 0.05). However, the addition of protease in the NC diet reinstated measures of intestinal function, as NC + PROT-fed piglets showed similar duodenal villus height, crude protein digestibility, gene expressions of duodenal peptide transporter PepT1 and jejunal amino acid transporter b0,+AT, as well as blood urea nitrogen and diamine oxidase concentrations to PC. In conclusion, dietary supplementation of protease in weaned piglets fed higher levels of lower digestible protein sources mitigated losses in growth performance and intestinal functionality by enhancing intestinal development, protein digestibility, nutrient transport efficiency, and overall health status of piglets.

Effects of dietary protease supplementation on in vitro soybean meal protein, dry matter digestibility, and productive performance in starter-to-finisher pigs.

Pig industries are currently facing a crisis in terms of protein and energy costs. Proteases were used to increase protein digestibility and metabolizable energy (ME) in diets. This study evaluated the effects of protease supplementation on in vitro protein digestibility and productive performance in starter-to-finisher pigs. A total of 691 starter pigs were randomly allocated into three dietary treatments using a randomized complete block design. Diets were provided in three phases according to body weight (BW): Starter, grower, and finisher phases. Each phase was fed for 30, 60, and 24 days of treatment diets as T1: basal diet and T2 and T3: the basal diet supplemented with 240 ppm protease reduced by 50 kcal/kg ME plus 1% crude protein (CP) and by 100 kcal/kg ME plus 2% CP, respectively. Protease and in vitro protein digestibility were measured. BW and feed intake were recorded to calculate the average daily gain (ADG), average daily feed intake (ADFI), feed-to-gain (F:G), and gain-to-feed (G:F) ratios. There were no significant differences (p > 0.05) in the percentage of in vitro protein digestibility between the groups with and without protease supplementation. In the finisher phase, T2 had lower (p < 0.05) ADFI and F:G than T1 and T3. Overall, T3 had lower (p < 0.05) ADG, ADFI, and F:G than T1 and T2. Protease supplementation significantly affects protein digestibility. Supplementing basal diets with 240 ppm protease reduced ME by 50 kcal/kg and CP by 1% without affecting ADG, ADFI, F:G, and G:F ratios for starter-to-finisher pigs.

Dietary Protease Supplementation Improved Growth Performance and Nutrients Digestion via Modulating Intestine Barrier, Immunological Response, and Microbiota Composition in Weaned Piglets.

Despite mounting evidence for dietary protease benefits, the mechanisms beyond enhanced protein degradation are poorly understood. This study aims to thoroughly investigate the impact of protease addition on the growth performance, intestinal function, and microbial composition of weaned piglets. Ninety 28-day-old weaned pigs were randomly assigned to the following three experimental diets based on their initial body weight for a 28-day experiment: (1) control (CC), a basic diet with composite enzymes without protease; (2) negative control (NC), a diet with no enzymes; and (3) dietary protease (PR), a control diet with protease. The results show that dietary proteases significantly enhanced growth performance and boosted antioxidant capacity, increasing the total antioxidant capacity (T-AOC) levels (p < 0.05) while reducing malonaldehyde levels (p < 0.05). Additionally, protease addition reduced serum levels of inflammatory markers TNF-α, IL-1β, and IL-6 (p < 0.05), suppressed mRNA expression of pro-inflammatory factors in the jejunum (p < 0.01), and inhibited MAPK and NF-κB signaling pathways. Moreover, protease-supplemented diets improved intestinal morphology and barrier integrity, including zonula occludens protein 1(ZO-1), Occludin, and Claudin-1 (p < 0.05). Microbiota compositions were also significantly altered by protease addition with increased abundance of beneficial bacteria (Lachnospiraceae_AC2044_group and Prevotellaceae_UCG-001) (p < 0.05) and reduced harmful Terrisporobacter (p < 0.05). Further correlation analysis revealed a positive link between beneficial bacteria and growth performance and a negative association with inflammatory factors and intestinal permeability. In summary, dietary protease addition enhanced growth performance in weaned piglets, beneficial effects which were associated with improved intestinal barrier integrity, immunological response, and microbiota composition.

PSII-17 Effect of protease supplementation on amino acids digestibility of soybean meal fed to growing pigs of different ages

Abstract Protease supplementation can increase amino acids (AA) availability; however, the effectiveness of protease in swine diets varies and has been associated with the type and dose of protease, dietary ingredients, and potentially animal age. This study investigated the interactive effect of protease inclusion level and pig age on standardized ileal digestibility (SID) of protein (CP) and AA of soybean meal (SBM) fed to growing pigs. A corn starch–based diet was formulated with SBM as the sole source of AA and containing titanium as an indigestible marker. Protease was supplemented at 0, 15,000; 30,000 and 45,000 NFP/kg of feed (ProAct 360). An N-free diet was used to estimate basal ileal endogenous AA losses. Ten cannulated pigs (body weight = 21 kg ± 2 kg) were assigned to experimental diets in a duplicate 5 × 5 Latin square design. In Phase I (23 to 30 kg-pigs, 90 ± 17 d of age), ileal digesta was collected in 5 periods of 7 d (5 d adaptation and 2 d ileal digesta collection). In Phase II (50 to 65 kg-pigs. 140 ± 17 d of age), using the same diets, ileal digesta was collected in 5 more periods of 7 d. Pigs were fed at 4% of body weight (BW) adjusted at the end of each period. Diets and ileal digesta were analyzed for CP, AA, and titanium and SID was calculated. Orthogonal polynomial contrasts were used to determine the linear and quadratic effects of protease supplementation. The combined data were analyzed as 4 x 2 factorial, including the fixed effects of protease inclusion (excluding N-free diet), age, and their interactions. In phase I, increasing levels of protease resulted in linear improved (P &amp;lt; 0.1) of SID for the majority of indispensable AA (Except Arg, His, Met + Cys, and Trp) and the average of dispensable AA. In phase II, the SID of Ile, Leu, Met, Met + Cys, Val, the average of indispensable AA, and the majority of dispensable AA was quadratically improved. Overall (Phase I and II), the SID of Ile, Leu, Met + Cys, Val, Cys, Glu, Tyr, and the average of dispensable AA was quadratically improved (P &amp;lt; 0.1). In most cases, the supplementation of 30,000 NFP/kg of feed resulted in the greatest AA digestibility improvements. However, the linear response in the first phase for some AA suggests that diets for younger pigs could be supplemented with a greater level (45,000 NFP/kg of feed). There were no interactions between protease levels and age. Younger pigs had improved (P &amp;lt; 0.1) SID of CP and most AA (Except Cys, Gly, and Ser). In conclusion, dietary protease supplementation can improve SID of AA in soybean mean independent of pig age. Pigs age can influence AA digestibility where younger pigs have greater AA digestibility.

Effect of a novel alkaline protease from Bacillus licheniformis on growth performance, carcass characteristics, meat quality, antioxidant capacity, and intestinal morphology of white feather broilers.

The present study was conducted to investigate the effects of dietary novel alkaline protease from Bacillus licheniformis on the growth performance, meat quality, antioxidant status and intestinal morphology of broilers. In total, 4000 broilers were randomly assigned into five groups and treated with normal control, normal control + 100 mg kg-1 protease, normal control + 200 mg kg-1 protease, normal control + 300 mg kg-1 protease and normal control + 400 mg kg-1 protease. Supplementing protease impacted final body weight (linear, P = 0.003; quadratic, P = 0.006) and decreased feed conversion rate (linear, P = 0.036) in broilers. Moreover, dietary protease significantly increased breast muscle rate (linear, P = 0.005; quadratic, P = 0.021) and decreased drip loss (linear, P < 0.001; quadratic, P < 0.001). In addition, dietary protease notably increased protein digestibility (linear, P = 0.001; quadratic, P = 0.006) and trypsin activity (linear, P = 0.002; quadratic, P = 0.009) in jejunum. Light microscopy revealed that the jejunum villi in the 300 mg kg-1 and 400 mg kg-1 groups exhibited greater height and a denser arrangement compared to those in the control group. The addition of protease decreased malondialdehyde content (linear, P < 0.001; quadratic, P < 0.001) and increased total antioxidant capacity (linear, P = 0.001; quadratic, P < 0.001) in pectoral muscles. The results of the present study suggest that dietary novel alkaline protease from B. licheniformis improved growth performance by affecting trypsin activity, protein digestibility, antioxidant capacity and intestinal health. © 2024 Society of Chemical Industry.

Diabrotica v. virgifera Seems Not Affected by Entomotoxic Protease Inhibitors from Higher Fungi.

Certain soil insects, such as the root-damaging larvae of the maize pest Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), are increasingly difficult to control because of recent bans of some insecticides. An alternative and safer approach may be the development of biopesticides based on entomotoxic defense proteins of higher fungi. Many of these potentially interesting proteins are protease inhibitors, and some have been shown to adversely affect insects. We examined the effects of the cysteine protease inhibitors macrocypin 1, 3, and 4 from Macrolepiota procera, clitocypin from Clitocybe nebularis, and cocaprin 1 and the serine protease inhibitor cospin 1 from Coprinopsis cinerea on D. v. virgifera. We confirmed the inhibition by mycocypins of the cysteine catalytic-type proteolytic activities in gut extracts of larvae and adults. The inhibition of pGlu-Phe-Leu-hydrolyzing activity was stronger than that of Z-Phe-Arg-hydrolyzing activity. Mycocypins and cospin resisted long-term proteolytic digestion, whereas cocaprin 1 was digested. Bioassays with overlaid artificial diet revealed no effects of proteins on neonatal mortality or stunting, and no effects on adult mortality. Immersion of eggs in protein solutions had little effect on egg hatching or mortality of hatching neonates. Microscopic analysis of the peritrophic matrix and apical surface of the midguts revealed the similarity between larvae of D. v. virgifera and the chrysomelid Leptinotarsa decemlineata, which are sensitive to these inhibitors. The resistance of D. v. virgifera to fungal protease inhibitors is likely due to effective adaptation of digestive enzyme expression to dietary protease inhibitors. We continue to study unique protein complexes of higher fungi for the development of new approaches to pest control.

PSV-15 Effects of Supplementation of Protease to Different Nutrient Density of Diets on Growth Performance, Nutrient Digestibility, Blood Profile, Gas Emission, Fecal Microflora in Growing-Finishing Pig

Abstract This study was conducted to investigate the effects of protease supplementation and different nutrient density of diets on growth performance, nutrient digestibility, blood profiles, gas emissions, and fecal microflora in growing-finishing pigs. Crossbred growing pigs (n = 180; [Landrace × Yorkshire] x Duroc) with an initial body weight (BW; 18.74 ± 3.46 kg) were used for 15 weeks. Pigs were randomly assigned to 6 dietary treatments with 6 replicates of 3 pigs per pen in a 3 ⅹ 2 factorial through the following arrangement: Three groups of protease (1, Basal diets; 2, Protease A: 125 mg/kg protease derived from Streptomyces spp; 3, Protease B: 100 mg/kg protease derived from Bacillus licheniformis) at two different nutrient density diets (1; Basal requirement 2; 0.94-0.98% greater than requirement in dietary protein and 50 kcal/kg in energy). Supplementation of protease significantly increased (P &amp;lt; 0.05) BW at weeks 4 and 6. Also, supplementation of protease increased (P &amp;lt; 0.05) ADG at grower periods and weeks 13 to 15. In addition, improvement (P &amp;lt; 0.05) of ADG was observed in pigs fed a high nutrient density diet (HN) compared with basal nutrient density diet (BN) at grower periods and weeks 13 to 15. In apparent total track digestibility (ATTD) of crude protein (CP), there was increased (P &amp;lt; 0.05) in supplementation of protease at weeks 6 and 13. The HN showed increased (P &amp;lt; .0001) ATTD of CP at 6 weeks. BN increased (P &amp;lt; 0.01) ATTD of CP at week 13. Also, there was interaction (P &amp;lt; 0.05) between nutrient density of diets and supplementation of protease. In ATTD of gross energy, supplementation of protease showed increased (P &amp;lt; 0.05) at week 13. The supplementation of protease in diets decreased (P &amp;lt; 0.05) the concentration of blood urea nitrogen (BUN) at week 13. The supplementation of protease in diets decreased (P &amp;lt; 0.05) the emission of NH3 at weeks 6 and 13. Also, H2S was decreased (P &amp;lt; 0.05) due to supplementation of Protease A in diets. At weeks 6 to 9, average daily feed intake showed a tendency to increase (P &amp;lt; 0.10) when the BN diets were fed. Also, during the entire period, feed conversion ratio tended to increase (P &amp;lt; 0.10) when the BN diets were fed. In conclusion, supplementation of protease reduces NH3 in feces and BUN in whole blood by increasing the digestibility of CP and improved growth performance. Also, diets with high nutrient density improved growth performance and nutrient digestibility in grower periods.

Exogenous protease influences protein digestibility, growth performance, and gut microflora in weanling pigs on a limited protein diet

ABSTRACTThe study was conducted to evaluate the impact of dietary level of crude protein (CP) and protease supplementation on growth performance, digestibility of nutrients, intestinal morphology, and gut microbiota in weaning pigs. Three hundred cross-bred piglets (Duroc × Landrace × Yorkshire) were allotted to five dietary treatments on the basis of initial body weight (BW) and sex. Pigs were group-housed in pens with each treatment with 10 replicate pens with six pigs per pen. The treatments included a standard diet (STD), STD with 0.6% lower protein (STD0.6), STD with 0.6% lower protein and protease supplementation (Pro0.6), STD with 1.0% lower protein (STD1.0), STD with 1.0% lower protein and protease supplementation (Pro1.0). Results indicated a higher BW (p < 0.05) of piglets in the Pro0.6 group at days 0–42 compared to the STD0.6 and STD1.0 groups. The average daily gain was higher (p < 0.05) in the Pro0.6 treatments at days 0–42 compared to the STD0.6 and STD1.0. The gain to feed ratio was higher (p < 0.05) in the STD, and Pro0.6 groups compared to the STD0.6, Pro1.0 and the STD1.0 groups at days 0–42. Dry matter digestibility was lower (p < 0.05) in the STD1.0 group than the Pro0.6 and Pro1.0 groups. The crude protein digestibility was higher (p < 0.05) in the Pro0.6 group compared to the STD, STD0.6 and STD1.0 treatment groups while crude fat digestibility was higher (p < 0.05) in the STD and Pro0.6 compared with the STD0.6 and STD1.0 groups. Digestibility was higher for histidine (p < 0.05), leucine (p < 0.05) in the protease Pro0.6 and Pro1.0 groups than in the STD0.6 and STD1.0 groups. The digestibility of non-essential AA was higher for alanine (p < 0.05) in the Pro0.6 than the STD1.0 group. For faecal microbial population, Faecalibacterium abundance was higher (p < 0.05) in the Pro0.6 compared to all the other groups while the population of Actinobacteria was greater (p < 0.05) in the STD group and lowest in the Pro1.0 treatment. In the ileum, villus height was greater (p < 0.05) in the protease Pro0.6, and Pro1.0 groups compared to the STD0.6, and STD1.0 groups while the villus height to crypts depth ratio was lower (p < 0.05) in the STD 1.0 group compared to the STD, Pro0.6, and Pro1.0 groups. Based on these results, dietary protease supplementation improved nutrient digestibility and gut histo-morphology translating to improved utilisation of nutrients thus positively impacting growth performance in weaned pigs. Further, reducing the CP content in the diets increased the abundance of Muribaculaceae while protease supplementation increased the population of Faecalibacterium in the gut of the weanling piglets on the STD0.6 diet.

Effects of dietary protease supplementation on behaviour, slaughter performance, meat quality and immune organ indices of broilers

Effects of dietary protease supplementation on behaviour, slaughter performance, meat quality and immune organ indices of broilers

Effects of Dietary Crude Protein and Protease Levels on Performance, Immunity Capacity, and AA Digestibility of Broilers

Exogenous proteases are promising to stimulate the application of low-protein diets for broilers. A total of 540 1-day-old Arbor Acres male broilers were randomly assigned to 9 groups with 6 replicates of 10 birds. A 3 × 3 factorial, completely randomized arrangement was used to evaluate the effects of dietary crude protein (CP) and protease levels on growth and slaughter performance, immunity capacity, and apparent ileal digestibility (AID) of amino acids (AA). Dietary CP levels were 20.0%, 19.5%, or 19.0% during the starter phase, and 18.0%, 17.5%, or 17.0% during the finisher phase. Protease levels were 0, 250, or 500 mg/kg in diets throughout the trial. The trial lasted for 42 days. Weight gain and feed efficiency of broilers decreased as dietary CP lowered, but improved with protease supplementation. Dietary CP and protease levels had few effects and interactions on carcass characteristics, immune organ indexes, and immunoglobulin concentrations. The AID of most AA was improved by dietary CP decrease or protease supplementation. In conclusion, reducing dietary CP decreased the performance and immune capacity of broilers but increased the AID of AA. Almost independent of dietary CP level, dietary protease addition improved the performance of broilers, probably through the enhancement of AA digestibility, and had no effect on carcass traits.

Influence of Dietary Protease on Growth Performance and Carcass Yield of Indian River Meat Broiler Chickens

The aim of the present study was to evaluate the effect of a multi-component dietary protease complex on the growth performance and carcass yields of commercial broiler chickens. A total of 630 Indian River broiler DOCs were considered and assigned to five dietary treatments namely: 1) Positive control (PC), a standard diet with no supplemental protease, 2) Negative control (NC), a diet similar to PC but reduced in CP, digestible EAA, and ME based on supplier’s nutrient matrix recommendation, 3) PC+125 ppm protease (PC+125), 4) NC+125 ppm protease (NC+125), and 5) NC+200 ppm protease (NC+200). The birds were fed experimental diets for 28 days. Body weight (BW), body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), and survival rate were recorded weekly. On day 28, three birds per treatment were sacrificed for carcass yield analysis. FI was similar among the treatment groups except in the 2nd week where broilers fed PC+125, NC+125, and NC+200 diets had significantly higher (P&lt;0.05) FI than those fed the PC diet. In terms of BW and BWG, there were no differences among the treatment groups at 1st and 2nd weeks, however, at 3rd and 4th weeks, broilers fed the PC+125, NC+125, and NC+200 diets were significantly heavier (P&lt;0.05) than those fed the PC and NC diets. A similar trend was also observed in FCR. Among the carcass yield parameters, the percentage weights of thigh, breast, and heart in birds fed the NC+200 diet were significantly increased (P&lt;0.05) as compared to those fed the NC diet, while results for PC, PC+125, and NC+125 were intermediate between the two treatment groups. Overall, the results of present study demonstrated that the supplementation of protease complex either on a standard or reduced diet may improve the overall production performance of commercial broiler chickens and offers benefits on certain aspects of carcass yield.

Effects of broiler chicken age and dietary protease on the standardised ileal digestibility of amino acids in seeds from two lupin species

Abstract The extent to which lupin seeds can replace soybean meal in diets for broiler chickens is limited, and one of the reasons for this may be a lack of careful consideration given to the age-related amino acid availability differences in the feed formulation process. This study aimed to determine and compare standardised ileal digestibility (SID) coefficients of amino acids (AA) in raw seeds of the Lupinus luteus (YL) and the Lupinus angustifolius (NLL) cultivars as sole sources of AA in the diet of broiler chickens aged 14 or 28 days. An additional purpose was to check the influence of exogenous mono-component protease added to lupin-based assay diets on AA SID in birds at both ages. Each assay diet was offered to six replicatecages. On both sampling days, the contents from the lower half of the ileum were collected for determination of the apparent digestibility values. The SID coefficients were calculated using the age-appropriate basal ileal endogenous AA losses determined from birds fed a N-free diet. Results indicated a substantial advantage of YL over the NLL in SID of Met, Cys, His, Leu, Gly, Asp, Glu, Pro, Tyr and the average of total AA for 14-d-old chickens. With the exception of Cys and Tyr, there was no significant difference between these Lupinus species in the SID of AA in 28-d-old birds. The significant age-related differences within species were found only for the seeds of NLL, where the higher SID for Met, His, Ala, Pro, and the averages of total and indispensable AA were noted in birds aged 28 d. The protease increased SID of most AA from L. luteus in older broilers, whereas for L. angustifolius its efficacy was more pronounced in younger birds, giving the SID values similar to those determined at 28 d on an enzyme-unsupplemented diet. In conclusion, the SID coefficients of indispensable AA determined at 14 or 28 days of age for yellow lupin should not be applied to the precise formulation of starter-type feeds containing seeds of NLL.

Effects of protease enzyme supplementation in weanling pigs' diet with different crude protein levels on growth performance and nutrient digestibility.

The purpose of this study is to determine the impact of different levels of crude protein (CP) diets supplemented with dietary protease on the growth performance and nutrient digestibility of the weanling pigs. In a 5-week study, 100 crossbred ([Landrace × Yorkshire] × Duroc) of weaner pigs that have an average initial body weight (BW) of 7.17±1.06 kg were assigned to one of four dietary treatments with 5 replications and 5 pigs (3 gilts and 2 castrated male pigs) per pen in a randomized complete block design. The dietary treatments were as follows: Phase 1: CON: basal diets (20.60% CP); low protein (LP): CON − 0.30% CP; PLP1: (CON − 0.30% CP) + 0.05% protease; PLP2: (CON − 0.50% CP) + 0.05% protease. Phase 2: CON: basal diets (18.88% CP); LP: CON − 0.30% CP; PLP1: (CON − 0.30% CP) + 0.05% protease; PLP2: (CON − 0.50% CP) + 0.05% protease. The addition of protease to low CP diets significantly increased the feed conversion ratio (FCR) (p = 0.039), BW (p = 0.046), average daily gain (ADG) (p = 0.049), and average daily feed intake (ADFI) tended to increase (p = 0.053) in the young pigs during phase 1. However, FCR tended to increase throughout the experiment but did not change during phase 2, whereas BW, ADG, and ADFI stayed unchanged throughout phase 2 and overall. There was no significant difference in dry matter, nitrogen (N), and gross energy of nutrient digestibility in all phases and overall in weaned pigs with low CP when protease was fed. In contrast, adding protease to the low CP diets increased the tendency of N digestibility (p = 0.059) during phase 1. It is concluded that dietary protease supplementation tended to increase N retention during the first phase of the weaning period, hence increasing piglet performance.

Modification of Gut Microbiota and Immune Responses via Dietary Protease in Soybean Meal-Based Protein Diets.

Plant-based protein sources such as soybean meal have low digestibility and are generally promoted accumulation of undigested proteins into the intestine by enzymatic treatments. Moreover, potential intestinal pathogens ferment undigested proteins, producing harmful substances, such as ammonia, amines and phenols, leading to an overactive immune response and diarrhea in weaned pigs. As a solution, dietary proteases hydrolyze soybean-based antinutritive factors, which negatively affect immune responses and gut microbiota. In this study, we investigated the effects of dietary proteases (PRO) in a low-crude protein (CP) commercial diet on the immune responses and gut microbiota of weaned pigs. The experimental design consisted of three dietary treatments: a commercial diet as a positive control (PC; phase1 CP = 23.71%; phase 2 CP: 22.36%), a lower CP diet than PC as negative control (NC; 0.61% less CP than PC), and NC diet supplement with 0.02% PRO. We found that PRO tended to decrease the frequency of diarrhea in the first two weeks after weaning compared with PC and NC. In addition, pigs fed PRO showed decreased TNF-α and TGF-β1 levels compared with those fed PC and NC. The PRO group had a higher relative proportion of the genus Lactobacillus and lower levels of the genus Streptococcus than the PC and NC groups. In conclusion, the addition of PRO to a low CP commercial weaned diet attenuated inflammatory responses and modified gut microbiota in weaned pigs.

The effectiveness of protease supplemented poultry by‐product meal‐based diet on growth, nutrient digestibility and digestive enzyme activities of rohu ( Labeo rohita )

Proteases are the enzymes responsible for the hydrolysis of complex proteins into simpler units for better digestion and absorption of nutrients. This study was conducted to determine the optimum supplementation level and effects of dietary protease addition in poultry by-product meal-based diet (PBM) on growth performance, nutrient digestibility, digestive enzyme activities and physiochemical parameters of juvenile rohu, Labeo rohita (initial weight: 11.33 ± 0.02 g). For this purpose, seven iso-nitrogenous and iso-lipidic experimental diets were formulated to contain a positive control (PC) diet (containing fishmeal and no protease), a negative control (NC) diet (containing PBM and no protease) and five other PBM-based diets supplemented with exogenous protease at the levels of 150, 300, 450, 600 and 750 mg/kg diet. Results indicated that growth performance and body indices of fish fed protease supplemented diets were higher (p < 0.05) compared with those fed the NC diet, while maximum increase was recorded at 450 mg/kg protease supplementation level, which was similar to the PC diet (p > 0.05). Protease supplementation up to 450 mg/kg level linearly improved (p < 0.05) the whole-body proximate composition and digestibility of dry matter (DM), crude protein (CP), crude lipid (CL) and crude ash (CA) in rohu juveniles. A similar trend (p < 0.05) was observed in the activities of amylases, lipases and proteases in intestinal and hepatopancreatic tissues, peaking at 450 mg/kg which was also similar (p > 0.05) to the PC diet. Protease supplementation significantly improved (p < 0.05) haemoglobin (Hb), haematocrit (Htc) and red blood cells (RBCs) of juveniles while, white blood cells (WBCs) and serum indices remained unaffected (p > 0.05). Based on quadratic regression analysis, the optimal dietary protease supplementation in PBM-based diet for rohu was 433 mg/kg. Conclusively, increase in protease supplementation from 0 to 450 mg/kg levels improved the fish performance and decreased thereafter with further increase in protease levels (600–750 mg/kg).

Effects of dietary protease supplementation on growth rate, nutrient digestibility, and intestinal morphology of weaned pigs.

The addition of dietary proteases (PRO) to weaner diets hydrolyzes soybean-based anti-nutritive factors and improves weaned pig’s dietary digestibility and growth performance. Therefore, this study explores the effects of PRO in a lower crude protein (CP) level diet than that in a commercial diet on the growth performance, nutrient digestibility, and intestinal morphology of weaned pigs. A total of 90 weaned pigs were randomly assigned to 3 dietary treatments with 6 pigs per pen and 5 replicated pens per treatment using a randomized complete block design (block = body weight [BW]): 1) a commercial weaner diet as a positive control (PC; phase1 CP = 23.71%; phase2 CP: 22.36%), 2) lower CP diet than PC as a negative control (NC; 0.61% less CP than PC), and 3) an NC diet with 0.02% PRO. Pigs fed PC and PRO had higher (p < 0.05) final BW, average daily gain, and/or gain to feed ratio for the first three weeks and the overall experimental period than NC. The PC and PRO groups had greater (p < 0.05) apparent ileal digestibility of dry matter, CP, and energy than the NC group. Moreover, pigs fed PC and PRO increased (p < 0.05) apparent total tract digestibility of CP compared with those fed NC. In addition, the PRO group had a higher number of goblet cells than the PC and NC groups. However, pig fed PC and PRO increased (p < 0.05) villus height and height to crypt depth ratio in the ileum compared with those fed NC. In conclusion, PRO supplementation in a commercial weaner diet with low CP levels improves growth rate and nutrient digestibility by modulating the intestinal morphology of weaned pigs.

Effect of supplementation of acid protease in diets containing different levels of crude protein on performance of white leghorn layers

Effect of supplementation of acid protease in diets containing different levels of crude protein on performance of white leghorn layers

The effects of dietary protease enzyme on growth performances and digestive enzyme activities of gilt head sea bream (Sparus aurata)

The effects of dietary protease enzyme on growth performances and digestive enzyme activities of gilt head sea bream (Sparus aurata)