Effects of different feeding regimens with protease supplementation on growth, amino acid digestibility, economic efficiency, blood biochemical parameters, and intestinal histology in broiler chickens.

BACKGROUND: This study was conducted to estimate the impacts of using varied feeding regimens with or without protease supplementation on the growth performance, apparent amino acid ileal digestibility (AID%), economic efficiency, intestinal histology, and blood biochemical parameters of broiler chickens. Three hundred one-day-old chicks (Ross 308 broiler) were randomly allotted to a 3 × 2 factorial design. The experimental design consisted of three feeding regimens; FR1: a recommended protein SBM diet, FR2: a low-protein SBM diet, and FR3: a low-protein diet with the inclusion of 5% DDGS and 5% SFM, with or without protease supplementation (250 mg/kg). RESULTS: Increased feed intake and feed conversion ratio were observed in the FR3 treatment during the starter stage and decreased body weight and body weight gain during the grower stage. However, there was no significant effect of the different feeding regimens, protease supplementation, or interaction on the overall performance. The economic value of diets also remained unaffected by the different feeding regimens, protease supplementation, or interaction. Protease supplementation resulted in lowering the AID% of tryptophan and leucine. Reduced AID% of methionine was evident in the FR2 + VE and FR3 - VE treatments. Histological findings substantiated the FR3 treatment mediated a decrease in the duodenal and jejunal villous height (VH), jejunal villous width (VW), and ileal VW, whereas, increase in the ileal crypt depth (CD). The FR2 + VE treatment reduced the VH:CD ratio in the duodenum. The duodenal CD and the jejunal goblet cell count were reduced as a consequence of protease supplementation. The FR3 + VE treatment documented a rise in duodenal CD, while an increase in the jejunal goblet cell count was observed in the FR3 - VE treatment. The FR3 treatment enhanced the IgM serum levels compared to the FR1 and FR2 treatments. IgM serum levels were also elevated following protease supplementation. FR3 + VE treatment increased IgM serum levels. The highest serum ALP was found in the FR3 treatment, whereas the lowest level was obtained in the FR2 treatment. CONCLUSION: Low-protein SBM-based diets could be used without affecting the birds' growth. Altered morphometric measures of the intestine and increased IgM and ALP levels indicated the low-protein SBM/DDGS-SFM diet-induced damage of the intestinal histoarchitecture and immune system of birds. These different diets and protease supplementation failed to affect economic efficiency positively.

Combined effects of enzymes and probiotics on hemato-biochemical parameters and immunological responses of juvenile Siberian sturgeon (Acipenser baerii).

A 10-week feeding trial was run to investigate the separate and simultaneous effects of exogenous enzymes (Enz), probiotics (Pro), and Pro-Enz mixtures on the hematology indices, serum biochemical parameters, and innate-immunity status of juvenile Siberian sturgeon. The fish (138.06 ± 3.64 g) were randomly dispersed into 12 tanks (20 individuals per tank) and fed with Enz (Phytase, protease, and xylanase), Pro (Pediococcus pentosaceus and Lactococcus lactis), and Pro-Enz cocktail. At the end of the feeding bioassay, the highest values of red blood cell count, hemoglobin concentration, hematocrit level, and lymphocyte percentage followed by the lowest neutrophil percentage were obtained in Pro-Enz treatment (P < 0.05). Despite a significantly lower level of alkaline phosphatase in the fish fed with Pro supplemented diet (P < 0.05), no significant difference was found in the serum level of alanine aminotransferase and aspartate aminotransferase among the experimental groups (P > 0.05). Total protein content was significantly upregulated in serum and skin mucus samples from those fed with supplemented diets compared to the control group (P < 0.05). In both serum and skin mucus samples, higher immune responses in terms of lysozyme activity, immunoglobulin M, total protein was seen in Pro-Enz treatment compared to the control group followed by the serum complement components (P < 0.05). The results indicate that the combinational supplementation of Siberian sturgeon diet with the exogenous enzymes and probiotics modulates the physiometabolic responses and innate immune system to a higher grade than their individual supplementation.

Interactive effect of exogenous protease enzyme and dietary crude protein levels on growth and digestibility indices in broiler chickens during the starter phase.

This study was conducted to study the effect of protease as well as three dietary crude protein levels on growth and nutrient digestibility in broilers. The broiler chickens (n = 540) were assigned to six dietary treatments with six replicates. Three dietary crude protein (CP) levels (17, 19, and 21%) along with two levels of protease (0 and 30,000 IU/kg) were investigated in 3 × 2 factorial arrangement during the starter phase. Body weight was higher (P < 0.05) in group fed with CP-21; however, feed intake was significantly (P < 0.05) lower in CP-19 and CP-21. Resultantly, FCR was significantly (P < 0.05) improved in birds fed with CP-21. Similarly, body weight, feed intake, and FCR were significantly (P < 0.05) improved in enzyme-treated birds compared to the untreated birds. For practical purpose, birds fed CP-19 with protease enzyme resulted in improved FCR compared to birds fed with CP-21. The results revealed that CP and apparent metabolizable energy (AME) were significantly (P < 0.05) higher in birds fed with CP-21 along with supplementation of enzyme while abdominal fat was significantly (P < 0.05) lower in the same group compared to birds fed with CP-17. These results indicated that a diet having 21% CP supplemented with 30,000 IU/kg protease enzyme was superior in improving the performance and digestibility of nutrients in broiler during the starter phase.

Daily Oral Administration of Protease-Treated Royal Jelly Protects Against Denervation-Induced Skeletal Muscle Atrophy.

Honeybees produce royal jelly (RJ) from their cephalic glands. Royal jelly is a source of nutrition for the queen honey bee throughout its lifespan and is also involved in fertility and longevity. Royal jelly has long been considered beneficial to human health. We recently observed that RJ delayed impairment of motor function during aging, affecting muscle fiber size. However, how RJ affects skeletal muscle metabolism and the functional component of RJ is as of yet unidentified. We demonstrate that feeding mice with RJ daily prevents a decrease in myofiber size following denervation without affecting total muscle weight. RJ did not affect atrophy-related genes but stimulated the expression of myogenesis-related genes, including IGF-1 and IGF receptor. Trans-10-hydroxy-2-decenoic acid (10H2DA) and 10-hydroxydecanoic acid (10HDAA), two major fatty acids contained in RJ. After ingestion, 10H2DA and 10HDAA are metabolized into 2-decenedioic acid (2DA) and sebacic acid (SA) respectively. We found that 10H2DA, 10HDAA, 2DA, and SA all regulated myogenesis of C2C12 cells, murine myoblast cells. These novel findings may be useful for potential preventative and therapeutic applications for muscle atrophy disease included in Sarcopenia, an age-related decline in skeletal muscle mass and strength.

Camelid VHH Antibodies that Neutralize Botulinum Neurotoxin Serotype E Intoxication or Protease Function.

Botulinum neurotoxin (BoNT) serotype E is one of three serotypes that cause the preponderance of human botulism cases and is a Tier 1 Select Agent. BoNT/E is unusual among BoNT serotypes for its rapid onset and short duration of intoxication. Here we report two large panels of unique, unrelated camelid single-domain antibodies (VHHs) that were selected for their ability to bind to BoNT/E holotoxin and/or to the BoNT/E light chain protease domain (LC/E). The 19 VHHs which bind to BoNT/E were characterized for their subunit specificity and 8 VHHs displayed the ability to neutralize BoNT/E intoxication of neurons. Heterodimer antitoxins consisting of two BoNT/E-neutralizing VHHs, including one heterodimer designed using structural information for simultaneous binding, were shown to protect mice against co-administered toxin challenges of up to 500 MIPLD50. The 22 unique VHHs which bind to LC/E were characterized for their binding properties and 9 displayed the ability to inhibit LC/E protease activity. Surprisingly, VHHs selected on plastic-coated LC/E were virtually unable to recognize soluble or captured LC/E while VHHs selected on captured LC/E were poorly able to recognize LC/E coated to a plastic surface. This panel of anti-LC/E VHHs offer insight into BoNT/E function, and some may have value as components of therapeutic antidotes that reverse paralysis following BoNT/E exposures.

Soybean meal allergenic protein degradation and gut health of piglets fed protease-supplemented diets.

Two experiments were conducted to determine the effects of protease supplementation on degradation of soybean meal (SBM) allergenic proteins (glycinin and ß-conglycinin) and gut health of weaned pigs fed soybean meal-based diets. In experiment 1, 2 SBM samples from 2 different sources were subjected to porcine in vitro gastric degradation to determine the effects of protease (at 15,000 U/kg of feedstuff) on degradation of the soybean allergenic proteins. In experiment 2, 48 weaned pigs (body weight = 6.66 kg) were obtained in 2 batches of 24 pigs each. Pigs were individually housed in metabolic crates and fed 4 diets (12 pigs/diet). The diets were corn-based diet with SBM 1 or SBM 2 without or with protease at 15,000 U/kg of diet in 2 × 2 factorial arrangement. Diets were fed for 10 d and pigs were sacrificed on day 10 for measurement of small intestinal histomorphology, permeability of small intestine mounted in Ussing chambers, and serum concentration of pro-inflammatory cytokines. Two SBM sources (SBM 1 and SBM 2) contained 46.9% or 47.7% CP, 14.0% or 14.6% glycinin, and 9.90% or 10.3% ß-conglycinin, respectively. Protease and SBM source did not interact on any of the response criteria measured in the current study. Protease supplementation tended to increase (P = 0.069) the in vitro gastric degradation of glycinin. Protease supplementation tended to reduce (P = 0.099) fluorescein isothiocyanate dextran 4,000 Da (which is a marker probe for intestinal permeability) flow in jejunum, and reduced (P = 0.037) serum TNF-α concentration. Protease did not affect small intestinal histomorphology. In conclusion, protease tended to increase gastric degradation of glycinin and reduce gut permeability, and serum concentration of pro-inflammatory cytokines, indicating that the protease used in the current study can be added to SBM-based diets for weanling pigs to improve gut health.

Protease em dietas com baixa proteína contendo farinha de Penas para codornas de corte / [Protease in low-protein diets for meat-type quails containing feather meal]

Objetivou-se avaliar a suplementação de protease em dietas com baixa proteína contendo farinha de penas (FP) sobre o desempenho e o rendimento de carcaça de codornas de corte. Foram utilizadas 240 codornas, machos, distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 2x 3 (com e sem protease x 3 níveis de FP (0%, 5% e 10%)), totalizando seis tratamentos (dieta reduzida (DR) em 8% da exigência de proteína bruta e aminoácidos + 0% FP; DR + 5% FP; DR + 10% FP; DR + 0% FP + protease; DR + 5% FP + protease e DR + 10% FP + protease), quatro repetições de 10 codornas por parcela, nas fases de oito-21 dias e oito-35 dias de idade. Observou-se interação (P≤0,05) entre os níveis de FP e protease no ganho de peso de oito-21 dias. Os níveis de FP influenciaram (P≤0,05) o consumo de ração e o ganho de peso de oito-21 e oito-35 e a conversão alimentar de oito-21 dias. Verificou-se interação (P≤0,05) entre aprotease e a inclusão de FPpara o peso corporalaos 35 dias. Conclui-se que aFP pode ser utilizada em até 5% em dietas para codornas de corte semsuplementação comprotease.(AU)

The objective of this study was to evaluate protease supplementation in low-protein diets containing feather meal (FP) on the performance and carcass yield of meat-type quails. Twenty male quails were used in a completely randomized design, in a 2 x 3 factorial scheme (with and without protease x 3 FP levels (0, 5 and 10%)), totaling six treatments (Reduced diet (RD) in 8 % of the requirement of crude protein and amino acids + 0% FP; RD + 5% FP; RD + 10% FP; RD + 0% FP + protease; RD + 5% FP + protease and RD + 10% FP + protease), four replicates of 10 quails per plot, in the phases of 8-21 days and 8-35 days of age. Interaction (P≤0.05) was observed between FP and protease levels on weight gain over the period of 8-21 days. The levels of FP influenced (P≤0.05) the feed intake and the weight gain of 8-21 and 8-35 and the feed conversion ratio of 8-21 days. There was interaction (P≤0.05) between protease supplementation and FP inclusion for body weight at 35 days. It is concluded that FP can be used up to 5% in diets for meat-type quails without protease supplementation.(AU)

Effects of dietary protein levels and protease supplementation on growth performance, carcass traits, meat quality, and standardized ileal digestibility of amino acid in Pekin ducks fed a complex diet.

This study aimed to investigate to the effects of dietary CP levels and protease supplementation on growth performance, carcass traits, meat quality, nutrients utilization, and standardized ileal digestibility of amino acid in Pekin ducks fed a complex diet. A total of 960 14-day-old male ducks were weighed and randomly allotted to a 2 × 5 factorial arrangement of 10 treatments with 6 replicate pens per treatment and 16 ducks per pen fed to 49 D of age. Experimental factors included five dietary CP levels ranging from 13.5 to 17.5% and with or without protease (200 mg/kg) supplementation. Between day 28 to 34, the digestible and metabolizable trials were performed. Significant CP × protease interactions (P < 0.05) on breast meat yield, DM, energy and nitrogen utilization, as well as standardized ileal digestibility values of 7 amino acids were observed. Regardless of protease supplementation, ducks fed 13.5, 14.5, and 15.5% CP had a poorer (P < 0.05) growth performance and breast meat yield than ducks fed with 16.5 and 17.5% CP. Ducks fed 13.5% CP had a positive effect (P < 0.05) on meat quality, dietary DM, energy and nitrogen utilization as well as standardized ileal digestibility of amino acids. Protease supplementation increased (P < 0.05) DM and phosphorus retention and decreased (P < 0.05) shear force of breast meat, regardless of CP level; when CP = 14.5%, protease significantly increased (P < 0.05) breast muscle yield. The optimal CP requirement without or with protease supplementation for BWG and FI were 17.02 or 16.53% and 16.64 or 16.75%, respectively, based on linear broken-line regression.

Toward standardized amino acid matrices for exogenous phytase and protease in corn-soybean meal-based diets for broilers.

A total of 468 male Ross 308 broilers were used in a digestibility study to determine the additivity of apparent or standardized amino acid (AA) digestibility values for corn, soybean meal (SBM), or a mixture of corn and SBM that were supplemented, or not, with either phytase, protease, or a combination of phytase and protease. These treatments generated a total of 12 experimental diets that were arranged in a 3 × 4 design. A nitrogen-free diet was also fed to estimate endogenous AA loss. Apparent and standardized AA digestibility values were assessed on day 28 posthatch. The apparent digestibility of AA in the complete diet was higher (P < 0.05) than expected based on the digestibility of the corn and SBM individually. However, this overestimation was corrected by the adjustment to standardized values. Importantly, addition of protease or the combination of protease and phytase increased (P < 0.05) the digestibility of AA in corn and SBM. Furthermore, these effects were arithmetically coherent with respect to the measured effects of the enzymes in the mixture of corn and SBM, even improving the additivity of AA digestibility values when assessed on an apparent basis. This study demonstrates that the effect of exogenous protease and phytase on AA digestibility in complete diets is predictable based on measurements made in individual ingredients. In addition to improving digestibility values per se, exogenous protease and phytase may enhance precision in least cost formulation systems.

Effects of intra-uterine infusion of proteolytic enzymes on selected cytokine concentrations, uterine inflammation, and fertility in postpartum water buffalo cows with subclinical endometritis.

The study was conducted to evaluate effects of intrauterine administration of proteolytic enzymes on endometrial inflammation and reproductive performance in postpartum water buffalo cows with subclinical endometritis (SCE). Cows (n = 38) with SCE (≥ 18 % PMN i.e.; polymorphonuclear cells) on day 21 postpartum (21 dpp), were allocated into treatment (TR; n = 19; intrauterine infusion of trypsin, chymotrypsin and papain in 20 ml normal saline on 21 dpp) and control (PC; n = 19; intrauterine administration 20 ml saline) groups. Cows without SCE (< 18 % PMN) were not treated and served as the negative control (NC; n = 30). Ultrasonography and sampling (endometrial cytology, uterine flushing, blood) were conducted on day 21 (before treatment) and 28 postpartum (28 dpp). The PMN % and uterine horn diameter were less on 28 dpp (compared with 21 dpp) in NC and TR group only. Cows with SCE had greater uterine concentrations of interleukin (IL)-1ß, IL-8 and tumor necrosis factor (TNF)-α; but lesser IL-10 than NC group on 21 and 28 dpp. There were greater serum IL-1ß and TNF-α concentrations on 28 dpp in cows with SCE than NC group. Uterine concentrations of IL-1ß were less, whereas IL-6 was greater following enzymatic treatment. Proteolytic enzyme treatment did not result in improvement in pregnancy rate compared with the PC group; however, days to conception were less in TR compared with the other two groups. In conclusion, results indicated a reduction in endometrial inflammation and days nonpregnant after proteolytic enzyme treatment in buffalo cows with SCE.

Influence of exogenous acid protease in broiler chickens fed corn-soybean meal-based diets.

This study was conducted to determine the influence of exogenous acid protease (EAP) on growth performance, blood profiles, excreta noxious gas emissions, ileum digesta viscosity and apparent ileal digestibility in broilers. A total of 600 1-d-old broilers with average initial body weight of 47.0 ± 0.2 g were used in a 5-week feeding trial. The broilers were randomly allotted to 1 of 4 treatments with 10 replicate pens per treatment and 15 birds per cage. Dietary treatments consisted of: CON, basal diets; EAP1, CON + 2,700 exogenous acid protease unite (APU)/kg; EAP2, CON + 5,400 APU/kg; EAP3, CON + 8,100 APU/kg. The body weight gain (BWG) had a linear improvement when chicks fed 8,100 AP/kg diets from d 18 to 35 and overall (p < .05). Lower feed conversion ratio (FCR) was obtained in broilers fed 5,400 APU/kg and 8,100 APU/kg as compared with the CON during overall (p < .05). Birds fed diets supplemented with EAP diet led to a linear increase in the ileal digestibility of crude protein, energy and amino acids (lysine, methionine, cystine, threonine, isoleucine, leucine, histidine and tryptophan) compared with CON group (p < .05). In conclusion, based on our results, we suggest that supplementation with EAP diet was effective in improving growth performance through increasing ileal digestibility of some nutrients (crude protein, energy and amino acids) in broilers.

Effect of protease derived from Pseudoalteromonas arctica supplementation on growth performance, nutrient digestibility, meat quality, noxious gas emission and blood profiles in finishing pigs.

This study was conducted to evaluate the effects of dietary supplementation of protease derived from Pseudoalteromonas arctica (PPA) in finishing pigs. A total of 160 pigs were used in this 10-week trial. Dietary treatment groups were as follows: CON (basal diet); TRT1 (basal diet + 0.1% PPA); TRT2 (basal diet + 0.2% PPA); and TRT3 (basal diet + 0.3% PPA). During weeks 1-5, pigs fed with different levels of PPA-supplemented diet showed linear increase (p < .05) in the apparent total tract digestibility (ATTD) of nitrogen (N) and linear decrease (p < .05) in the concentrations of serum total protein. During weeks 6-10, pigs fed with different levels of PPA-supplemented diet showed a linear decrease in feed conversion ratio (p < .05). During the overall period, there was a linear decrease in feed conversion ratio (p < .05) associated with the inclusion of PPA. Pigs fed diets with 0.2% PPA supplementation had lower (p < .05) feed conversion ratio than those fed CON diet during weeks 6-10 and the overall period, and had higher (p < .05) ATTD of N than those fed CON diet during weeks 1-5. Pigs fed diets with PPA supplementation had lower (p < .05) concentrations of serum total protein than those fed CON diet on week 5. In conclusion, dietary supplementation with PPA diet has beneficial effects on growth performance, nutrient digestibility, backfat thickness and the concentrations of serum total protein.

The potential of plant and fungal proteins in the control of gastrointestinal nematodes from animals.

Gastrointestinal nematode infection is an important cause of high economic losses in livestock production. Nematode control based on a synthetic chemical approach is considered unsustainable due to the increasing incidence of anthelmintic resistance. Control alternatives such as the use of natural products are therefore becoming relevant from an environmental and economic point of view. Proteins are macromolecules with various properties that can be obtained from a wide range of organisms, including plants and fungi. Proteins belonging to different classes have shown great potential for the control of nematodes. The action of proteins can occur at specific stages of the nematode life cycle, depending on the composition of the external layers of the nematode body and the active site of the protein. Advances in biotechnology have resulted in the emergence of numerous protein and peptide therapeutics; however, few have been discussed with a focus on the control of animal nematodes. Here, we discuss the use of exogenous proteins and peptides in the control of gastrointestinal.

The potential of plant and fungal proteins in the control of gastrointestinal nematodes from animals / Potencial de proteínas de plantas e fungos no controle de nematoides gastrintestinais de animais

Abstract Gastrointestinal nematode infection is an important cause of high economic losses in livestock production. Nematode control based on a synthetic chemical approach is considered unsustainable due to the increasing incidence of anthelmintic resistance. Control alternatives such as the use of natural products are therefore becoming relevant from an environmental and economic point of view. Proteins are macromolecules with various properties that can be obtained from a wide range of organisms, including plants and fungi. Proteins belonging to different classes have shown great potential for the control of nematodes. The action of proteins can occur at specific stages of the nematode life cycle, depending on the composition of the external layers of the nematode body and the active site of the protein. Advances in biotechnology have resulted in the emergence of numerous protein and peptide therapeutics; however, few have been discussed with a focus on the control of animal nematodes. Here, we discuss the use of exogenous proteins and peptides in the control of gastrointestinal.

Resumo A infecção por nematoides gastrintestinais é uma importante causa de grandes perdas econômicas na pecuária. O controle de nematoides com compostos químicos sintéticos é considerado insustentável devido ao aumento da resistência anti-helmíntica. Alternativas de controle, como o uso de produtos naturais, estão se tornando relevantes do ponto de vista ambiental e econômico. As proteínas são macromoléculas com várias propriedades que podem ser obtidas de uma ampla gama de organismos, incluindo plantas e fungos. Proteínas pertencentes a diferentes classes têm mostrado grande potencial para o controle de nematoides. A ação das proteínas pode ocorrer em estágios específicos do ciclo de vida do nematoide, dependendo da composição das camadas externas do parasito e do sítio ativo da proteína. Avanços na biotecnologia resultaram no surgimento de numerosas terapias de proteínas e peptídeos; no entanto, pouco foi discutido com foco no controle de nematoides parasitos de animais. Na presente revisão foi discutido o uso de proteínas exógenas e peptídeos no controle de nematoides gastrintestinais, os mecanismos sugeridos de ação, e os desafios e perspectivas para o uso dessas biomoléculas como uma classe de anti-helmínticos.

Prececal amino acid digestibility and phytate degradation in broiler chickens when using different oilseed meals, phytase and protease supplements in the feed.

The purpose of this study was to investigate the effects of phytase and protease supplementation on prececal (pc) amino acid (AA) digestibility, phytate (InsP6) degradation, and MEn concentration in diets using 3 oilseed meals as main protein sources in broiler chicken feed. The broiler chicken diets, which lacked mineral phosphorus, contained either soybean meal (SBM), SBM and rapeseed meal (SBM/RSM), or SBM and sunflower meal (SBM/SFM) as main protein sources. Diets were not supplemented with enzymes or supplemented with 1,500 or 3,000 FTU phytase/kg, or with 1,600 mg protease/kg. For diets containing SBM as the main protein source, the effects of phytase supplementation with and without monocalcium phosphate were also investigated. Data were obtained during 2 subsequent runs from days 14 to 22 and from days 23 to 31. Each diet was tested using 8 replicates with 4 replicates per run. For pc AA digestibility, no significant interactions were observed between main protein sources, enzyme supplementation, or addition of monocalcium phosphate except for Cys. Supplementation of 1,500 FTU phytase/kg increased pc digestibility of all AA. No differences in pc AA digestibility were observed between 1,500 and 3,000 FTU phytase/kg supplementation treatments. Prececal disappearance of InsP6 and pc P digestibility were greater in the high phytase supplementation treatment. Protease supplementation increased pc digestibility of all AA except for Cys when SBM/RSM was the main protein source. Supplementation of protease and 3,000 FTU phytase/kg increased MEn concentrations. The effect of phytase on pc AA digestibility was fully expressed at a lower supplementation level than needed for a maximized pc InsP6 disappearance and MEn concentration.

Effects of a combination of xylanase, amylase and protease, and probiotics on major nutrients including amino acids and non-starch polysaccharides utilization in broilers fed different level of fibers.

This study evaluated the effects of a combination of xylanase, amylase, and protease (XAP), with probiotics (3 Bacillus spp.) supplementation on apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients in Cobb 500 broilers from 0 to 21 d. A completely randomized 2 × 4 factorial design (2 levels of fiber; 4 types of supplements) with 8 replicate cages (6 birds/cage) was used. Each low and high-fiber diet contained 500 FTU/kg Buttiauxella sp. phytase and was supplemented with: (a) none (control), (b) XAP (2,000 U xylanase + 200 U amylase + 4,000 U protease/kg diet), (c) probiotics (75,000 CFU/g of Bacillus spp.), or (d) XAP + probiotics. High fiber decreased (P < 0.05) nitrogen-corrected apparent metabolizable energy (AMEn), AID of all amino acids (AA), AID and ATTD of dry matter (DM), crude protein (CP), starch, and gross energy (GE). High fiber increased (P < 0.01) the flow of total non-starch polysaccharides (NSP) in both ileum and total tract. The XAP + probiotics increased (P < 0.01) AMEn as well as AID and ATTD of DM, CP, GE, starch, while alone, XAP yielded similar improvement except for DM compared with control. The supplemental XAP alone improved (P < 0.01) the digestibility of most of the AAs compared with control. Moreover, XAP + probiotics increased (P < 0.05) AID of all AA except arginine and serine compared with control. A fiber × supplements interaction (P < 0.05) was found for AID of histidine and threonine, and their digestibility in high-fiber diet was improved to a level comparable to low-fiber diet by XAP + probiotics. The flow of NSP in XAP group was 5 to 6% lower than in control while NSP flow in XAP + probiotic group was further 4% lower than that of XAP group (P < 0.01). The results infer that the combination of XAP and probiotics can effectively optimize the nutrient digestibility in broilers fed both low and high-fiber diets.

Evaluation of novel protease enzymes on growth performance and nutrient digestibility of poultry: enzyme dose response.

Two experiments were conducted to evaluate 3 novel proteases in broilers. In experiment 1, 600 male, Cobb 500 broilers were allocated to 1 of 12 experimental diets (5 birds/pen and 10 replicates/diet). A control (C) diet was formulated to be adequate in all nutrients. Proteases 1, 2, or 3 were added to this diet at 3 doses (1x, 3x, or 9x) in a 3 × 3 factorial arrangement of treatments. The factorial was augmented with 2 treatments of phytase at 500 or 1,500 FTU/kg added to the C diet. In experiment 2, 2,050 male Ross 308 broilers were allocated to 1 of 10 experimental diets (25 birds/pen and 9 replicates/diet). A C diet was formulated to be adequate in all nutrients. Protease 1, 2, or 3 was then added to the C diet at 3 doses (1x, 2x, or 4x) in a 3 × 3 factorial arrangement of treatments plus the C. In experiment 1, birds fed phytase gained more (P < 0.05) than birds fed protease, but neither were different than birds fed the C. Supplementation of 9x dose of any protease resulted in a reduction (P < 0.05) in BWG when compared with birds fed 1x dose of protease or phytase at 500 or 1,500 FTU/kg. Feed conversion ratio was improved (P < 0.05) in birds fed phytase compared with birds fed the C diet. Nitrogen digestibility was greater (P < 0.05) in birds fed protease 1 when compared with birds fed protease 2. Birds fed the 1x dose of protease or 500 FTU/kg of phytase had a greater (P < 0.05) N digestibility than birds fed 3x dose of protease. In experiment 2, protease supplementation significantly reduced (P < 0.05) BWG when compared with birds fed the C from hatch to 35 D post-hatch. Protease supplementation did not improve broiler growth performance or N digestibility above that of a nutrient adequate control diet or a diet supplemented with 500 FTU/kg of phytase.

Feeding Aspergillus protease preparation combined with adequate protein diet to rats increases levels of cecum gut-protective amino acids, partially linked to Bifidobacterium and Lactobacillus.

Our recent study indicated that dietary Aspergillus oryzae-derived protease preparation (AP), through its enzymatic activity, exerted a bifidogenic effect in rats. We hypothesized that dietary AP links to protein degradation and subsequently elevates gut-protective amino acids (AAs) in rats fed adequate protein diet. In this study, dietary AP markedly increased the relative abundance of Bifidobacterium and Lactobacillus and the levels of free threonine, alanine, proline, taurine, ornithine, phenylalanine, cystine, and γ-aminobutyric acid in the cecum contents of rats fed with an adequate protein diet, but not in those fed with a low-protein diet. The elevated AAs, except ornithine and phenylalanine, potentially have gut-related health benefits. Some of the AP-modulated free AAs appeared to be associated with the relative abundance of Bifidobacterium and Lactobacillus. Thus, AP combined with adequate protein diet is likely to increase the levels of cecum beneficial free AAs, which is partially associated with the relative abundance of the probiotics.

Effect of acute exposure in swiss mice (Mus musculus) to a fibrinolytic protease produced by Mucor subtilissimus UCP 1262: An histomorphometric, genotoxic and cytological approach.

The fibrinolytic enzyme produced by Mucor subtilissimus UCP 1262 was obtained by solid fermentation and purified by ion exchange chromatography using DEAE-Sephadex A50. The enzyme toxicity was evaluated using mammalian cell lineages: HEK-293, J774.A1, Sarcoma-180 and PBMCs which appeared to be viable at a level of 80%. The biochemical parameters of the mice treated with an acute dose of enzyme (2000 mg/mL) identified alterations of AST and ALT and the histomorphometric analysis of the liver showed a loss of endothelial cells (P < 0.001). However, these changes are considered minimal to affirm that there was a significant degree of hepatotoxicity. The comet assay and the micronucleus test did not identify damage in the DNA of the erythrocytes of the animals treated. The protease did not degrade the Aα and Bß chains of human and bovine fibrinogens, thus indicating that it does not act as anticoagulant, but rather as a fibrinolytic agent. The assay performed to assess blood biocompatibility shows that at dose of 0.3-5 mg/mL the hemolytic grade is considered insignificant. Moreover, the enzyme did not prolong bleeding time in mice when dosed with 1 mg/kg. These results indicate that this enzyme produced is a potential competitor for developing novel antithrombotic drugs.

Nondietary Therapies for Celiac Disease.

Celiac disease (CD) is an autoimmune enteropathy triggered by gluten. Gluten-free diets can be challenging because of their restrictive nature, inadvertent cross-contaminations, and the high cost of gluten-free food. Novel nondietary therapies are at the preclinical stage, clinical trial phase, or have already been developed for other indications and are now being applied to CD. These therapies include enzymatic gluten degradation, binding and sequestration of gluten, restoration of epithelial tight junction barrier function, inhibition of tissue transglutaminase-mediated potentiation of gliadin oligopeptide immunogenicity or of human leukocyte antigen-mediated gliadin presentation, induction of tolerance to gluten, and antiinflammatory interventions.