Prediction of net energy of feeds for broiler chickens.

Net energy (NE) enables the prediction of more accurate feed energy values by taking into account the heat increment which is approximately 25% of apparent metabolizable energy (AME) in poultry. Nevertheless, application of NE in poultry industry has not been practiced widely. To predict the NE values of broiler diets, 23 diets were prepared by using 13 major ingredients (wheat, corn, paddy rice, broken rice, cassava pellets, full-fat soybean, soybean meal, canola meal, animal protein, rice bran, wheat bran, palm kernel meal and palm kernel oil). The diets were formulated in order to meet the birds' requirements and get a wide range of chemical compositions (on DM basis; 33.6% to 55.3% for starch; 20.8% to 28.4% for CP, 2.7% to 10.6% for ether extract [EE] and 7.0% to 17.2% for NDF), with low correlations between these nutrients and low correlations between the inclusion levels of ingredients allowing for the calculation of robust prediction equations of energy values of diets or ingredients. These diets were fed to Ross 308 broilers raised in 12 open-circuit respiratory chambers from 18 to 23 d of age (4 birds per cage) and growth performance, diet AME content and heat production were measured, and dietary NE values were calculated. The trial was conducted on a weekly basis with 12 diets measured each week (1 per chamber), 1 of the 23 diets (reference diet) being measured each week. Each diet was tested at least 8 times. In total, 235 energy balance data values were available for the final calculations. Growth performance, AME (15.3 MJ/kg DM on average) and AME/GE (79.4% on average) values were as expected. The NE/AME value averaged 76.6% and was negatively influenced by CP and NDF and positively by EE in connection with efficiencies of AME provided by CP, EE and starch for NE of 73%, 87% and 81%, respectively. The best prediction equation was: NE = (0.815 × AME) - (0.026 × CP) + (0.020 × EE) - (0.024 × NDF) with NE and AME as MJ/kg DM, and CP, EE and NDF as % of DM. The NE prediction equations from this study agree with other recently reported equations in poultry and are suitable for both ingredients and complete feeds.

Re-evaluation of recent research on metabolic utilization of energy in poultry: Recommendations for a net energy system for broilers.

Different energy systems have been proposed for energy evaluation of feeds for domestic animals. The oldest and most commonly used systems take into account the fecal energy loss to obtain digestible energy (DE), and fecal, urinary and fermentation gases energy losses to calculate metabolizable energy (ME). In the case of ruminants and pigs, the net energy (NE) system, which takes into account the heat increment associated with the metabolic utilization of ME, has progressively replaced the DE and ME systems over the last 50 years. For poultry, apparent ME (AME) is used exclusively and NE is not yet used widely. The present paper considers some important methodological points for measuring NE in poultry feeds and summarizes the available knowledge on NE systems for poultry. NE prediction equations based on a common analysis of three recent studies representing a total of 50 complete and balanced diets fed to broilers are proposed; these equations including the AME content and easily available chemical indicators have been validated on another set of 30 diets. The equations are applicable to both ingredients and complete diets. They rely primarily on an accurate and reliable AME value which then represents the first limiting predictor of NE value. Our analysis indicates that NE would be a better predictor of broiler performance than AME and that the hierarchy between feeds is dependent on the energy system with a higher energy value for fat and a lower energy value for protein in an NE system. Practical considerations for implementing such an NE system from the commonly used AME or AMEn (AME adjusted for zero nitrogen balance) systems are presented. In conclusion, there is sufficient information to allow the implementation of the NE concept in order to improve the accuracy of feed formulation in poultry.

Inorganic and Organic Selenium Speciation of Seleno-Yeasts Used as Feed Additives: New Insights from Elemental Selenium Determination.

Seleno-Yeasts (SY) used as feed additives are known to contain different Selenium (Se) species. Seleno-Yeasts has been shown, on previous analytical methods, to contain selenomethionine (SeMet), selenocysteine (SeCys), selenate (SeIV) and selenite (SeVI), and various other organic and inorganic Se forms identified but rarely quantified. A new advanced method has allowed elemental Se (Se0), an inorganic Se species, to be quantified, thereby obtaining better insight into the proportion of inorganic Se in SY products. The study aimed to quantify the Se0 in SY products and assess the proportion of inorganic Se in SY. The Se speciation of 13 fresh commercials SY from different suppliers and batches, was assayed for the total Se, inorganic Se species (SeIV, SeVI and Se0), and organic Se species (SeMet and SeCys). Results on total Se were in line with the expected Se concentrations for all evaluated samples. The proportion of Se present as Se0 ranged from 3.6% to 51.8%. The quantity of Se0 in the SY products, added to SeIV and SeVI, indicated an average proportion of inorganic Se of 14.2% for the 13 analyzed SY products. The proportion of Se as SeMet ranged from 19.0% to 71.8%, (average of 55.8%), and a large variability in the SeMet content was observed. The SeCys content was also variable, with an average of 3.8%, relative to the total Se. In conclusion, advances in the analytical characterization have revealed that SY products can have a significantly high proportion of inorganic Se, which could affect the bioavailability of Se from SY supplements and explain their variable and lower bio-efficacy than pure SeMet supplements, such as hydroxy-selenomethionine.

Organic Selenium (OH-MetSe) Effect on Whole Body Fatty Acids and Mx Gene Expression against Viral Infection in Gilthead Seabream (Sparus aurata) Juveniles.

The supplementation of fish diets with OH-SeMet reduces oxidative stress and modulates immune response against bacterial infection. However, despite the importance of essential polyunsaturated fatty acids in fish nutrition and their high risk of oxidation, the potential protective effect of OH-SeMet on these essential fatty acids has not been studied in detail. Moreover, while viral infection is very relevant in seabream production, no studies have focused the Se effects against viral infection. The aim of the present study was to assess the impact of dietary supplementation with OH-SeMet on gilthead seabream fatty acid profiles, growth performance and response against viral infection. Gilthead seabream juveniles (21.73 ± 0.27 g) were fed for 91 days with three experimental diets, a control diet without supplementation of Se (0.29 mg Se kg diet-1) and two diets supplemented with OH-SeMet (0.52 and 0.79 mg Se kg diet-1). A crowding stress test was performed at week 7 and an anti-viral response challenge were conducted at the end of the feeding trial. Selenium, proximate and fatty acid composition of diets and body tissues were analyzed. Although fish growth was not affected, elevation in dietary Se proportionally raised Se content in body tissues, increased lipid content in the whole body and promoted retention and synthesis of n-3 polyunsaturated fatty acids. Specifically, a net production of DHA was observed in those fish fed diets with a higher Se content. Additionally, both monounsaturated and saturated fatty acids were significantly reduced by the increase in dietary Se. Despite the elevation of dietary Se to 0.79 mg kg-1 not affecting basal cortisol levels, 2 h post-stress plasma cortisol levels were markedly increased. Finally, at 24 h post-stimulation, dietary OH-SeMet supplementation significantly increased the expression of the antiviral response myxovirus protein gene, showing, for the first time in gilthead seabream, the importance of dietary Se levels on antiviral defense.

Tissue localization of selenium of parental or dietary origin in rainbow trout (Oncorhynchus mykiss) fry using LA-ICP MS bioimaging.

In relation to the decrease of selenium (Se) content in aquafeeds, the impact of level and form of parental and dietary Se supplementation was investigated in rainbow trout fry using laser ablation-inductively coupled plasma mass spectrometry (LA-ICP MS) bioimaging. The offspring of rainbow trout broodstock, fed either a control diet without any Se supplementation (0.3 mg Se/kg diet) or a diet supplemented with Se (0.6 mg Se/kg diet) either as sodium selenite or hydroxy-selenomethionine, were sampled at swim-up fry stage or after 11 weeks of cross-feeding. Total body Se levels were influenced by parental Se nutrition in swim-up fry and by direct Se feeding in 11-week fry with higher levels in the Se-supplemented groups compared with the control and the highest levels in the hydroxy-selenomethionine treatment. The Se retention was lower for dietary sodium selenite. Selenomethionine levels increased when Se was provided as hydroxy-selenomethionine. LA-ICP MS maps revealed yolk in swim-up fry and intestine, liver, and kidney in 11-week fed fry as tissues with high Se abundance. In swim-up fry, muscle Se was the highest abundant when parents were fed hydroxy-selenomethionine. In 11-week fed fry, muscle Se abundance was higher in the head part of fry fed both Se-supplemented diets, but only in the tail part of fry fed hydroxy-selenomethionine. Liver Se abundance was higher in fry fed sodium selenite compared with the control diet supporting the hypothesis that tissue Se distribution can be influenced by parental and dietary Se forms and levels.

Sulphur amino acids, muscle redox status and meat quality: More than building blocks - Invited review.

Advances in food science ineludibly require a profound study of food chemistry fundamentals. Consequently, we need to explore the environment where life and all underlying biological processes occur: the molecular level. The progress made in the last few years in relation to meat protein biochemistry enables revisiting the role that sulphur amino acids (SAA) play as redox-active muscle components. Beyond their participation as building blocks in the de novo synthesis of muscle proteins, protein thiols and other sulphur-containing species are active in controlling metabolic pathways, enzymatic processes, and redox reactions. Supplementation of SAA over nutritional requirements has been found to protect muscle lipids and proteins against oxidation, with subsequent benefits in terms of water-holding capacity, colour stability and sensory properties. As strong nucleophilic compounds, protein thiols establish assorted molecular interactions with other meat and non-meat electrophilic species (i.e. phytochemicals). Those interactions have been found to display remarkable effects on the redox properties, functionality and digestibility of meat proteins. The comprehension of these chemistry fundamentals enables a rationale manipulation of SAA via animal nutrition and/or food formulation to guarantee meat quality and safety and furthermore, develop novel, safer and more nutritious muscle foods, with increased oxidative stability.

Reducing BW loss during lactation in sows: a meta-analysis on the use of a nonstarch polysaccharide-hydrolyzing enzyme supplement.

A meta-analysis was performed on eight trials, which included a total of 992 parity 1 to 8 lactating sows, to evaluate the effects of feeding xylanase which is the main enzyme activity present in the enzymatic complex (Rovabio Excel, Adisseo, France) supplement throughout lactation on the following sow performance factors: BW loss, feed intake, backfat depth, and piglet growth. Even a short period of enzyme supplementation during lactation led to a reduction in BW loss of approximately 3 kg per sow (P = 0.003). This reduction represented 1-2% of the BW of sows. This effect could be explained by an increase in feed energy intake and enhanced feed digestibility. Sows fed enzyme-supplemented diets exhibited greater DM, OM, and GE digestibilities (3.4, 3.9, and 4.2% increases, respectively; P < 0.001) than sows fed control diets. During lactation, sows lost from 19 to 25 kg of BW (i.e., approximately 10% of their BW), with a difference between parity groups (P < 0.001). Body reserve mobilization was decreased in sows fed enzyme-supplemented diets (-2.9 kg, P = 0.003), with a more pronounced effect in primiparous than multiparous sows when BW loss is expressed relative to total BW (-2.27 vs. -0.59%, respectively; P = 0.058). Enzyme supplementation also increased litter weight gain up to weaning, with a greater effect in litters from multiparous sows than those from primiparous sows (5.4 vs. 0.6 kg, respectively; P = 0.009). These results could be explained in part by the relationship between their NE intake and either variations in BW or litter weight gain (R2 = 0.51 and 0.49, respectively; P < 0.001). Finally, the meta-analysis suggests that there are differences in the partitioning of the NE intake between growth and milk production and in relation to the sow's parity or physiological status. Extra energy released by enzyme is used for one of these functions (i.e., body mobilization reduction or greater milk export for litter gain).

Next-generation non-starch polysaccharide-degrading, multi-carbohydrase complex rich in xylanase and arabinofuranosidase to enhance broiler feed digestibility.

This study was carried out to evaluate the effect of a multi-carbohydrase complex (MCC) rich in xylanase (Xyl) and arabinofuranosidase (Abf) on overall broiler feed digestibility in broilers. Energy utilization and digestibility of dry matter (DM), organic matter (OM), protein, starch, fat, and insoluble and soluble fibers were measured using the mass-balance method. The experiment was carried out on 120 broilers (3-week-old chickens). Broilers were distributed over 8 treatments to evaluate the effect of the dietary arabinoxylan content and nutrient density with and without MCC (Rovabio® Advance). The graded content of arabinoxylan (AX) was obtained using different raw materials (wheat, rye, barley, and dried distillers' wheat). Diet-energy density was modified with added fat. Measurements indicated that nutrient density and AX content had a significant effect on most digestibility parameters. Apparent metabolizable energy (AME) was significantly increased (265 kcal kg-1) by MCC. The addition of MCC also resulted in significant improvement in the digestibility of all evaluated nutrients, with average improvements of 3.0, 3.3, 3.2, 3.0, 6.2, 2.9, 5.8, and 3.8% units for DM, OM, protein, starch, fat, insoluble and soluble fibers, and energy utilization, respectively. The interaction between MCC and diet composition was significant for the digestibility of OM, fat, protein, and energy. Nutrient digestibility and diet AME were negatively correlated with AX content (P < 0.001). However, the addition of MCC resulted in a reduction of this negative effect (P < 0.001). The AME of diets with and without the addition of MCC were successfully predicted by the diet digestible nutrient (i.e., starch, protein, fat, insoluble and soluble fibers) content with and without MCC (R2 = 0.87; RSD = 78 kcal kg-1). This study confirms that the presence of AX in wheat-based diets and wheat-based diets with other cereals and cereal by-products reduces nutrient digestibility in broiler chickens. Furthermore, the dietary addition of MCC, which is rich in Xyn and Abf, reduced deleterious effect of fiber and improved overall nutrient digestibility in broiler diets.

Selenium deposition kinetics of different selenium sources in muscle and feathers of broilers.

The objective of this study was to determine selenium (Se) deposition kinetics in muscles and feathers of broilers in order to develop a rapid method to compare bioavailability of selenium sources. Different Se sources such as 2-hydroxy-4-methylselenobutanoic acid (HMSeBA, SO), sodium selenite (SS) and seleno-yeast (SY) were compared for their kinetics on Se deposition in muscles and feathers in broiler chicks from 0 to 21 d of age. A total of 576 day-old broilers were divided into four treatments with 8 replicates of 18 birds per pen. The diets used in the experiment were a negative control (NC) not supplemented with Se and 3 diets supplemented with 0.2 mg Se/kg as SS, SY or SO. Total Se content in breast muscle and feathers were assessed on days 0, 7, 14 and 21. At 7 d of age, SO increased muscle Se content compared to D0 (P < 0.05), whereas with the other treatments, muscle Se concentration decreased (P < 0.05). After 21 days, organic Se sources maintained (SY) or increased (SO) (P < 0.05) breast muscle Se concentration compared to hatch value whereas inorganic source (SS) or non-supplemented group (NC) showed a significant decrease in tissue Se concentration (P < 0.05). At D21, Se contents of muscle and feathers were highly correlated (R(2) = 0.927; P < 0.0001). To conclude, these results indicate that efficiency of different Se sources can be discriminated through a 7 d using muscle Se content in broiler chickens. Muscle and feathers Se contents were highly correlated after 21 days. Also feather sampling at 21 days of age represents a reliable and non-invasive procedure for Se bioefficacy comparison.

2-Hydroxy-4-methylselenobutanoic acid induces additional tissue selenium enrichment in broiler chickens compared with other selenium sources.

Two experiments were conducted in broiler chickens to compare the effect of different Se sources on Se tissue enrichment: sodium selenite (SS), seleno-yeast (SY), and a new organic Se source (SO) containing 2-hydroxy-4-methylselenobutanoic acid (HMSeBA) as an active substance. For each experiment, treatments differed only in source or dose of Se additive. Relative efficiency was compared by plasma and tissue [muscle (pectoralis major) and liver] total Se concentrations. The first experiment compared Se sources (SS, SY, and SO) at different concentrations (mg of Se/kg of feed; SS-0.3; SY-0.1 and -0.3; SO-0.1 and -0.3; and a negative control, 0) in broilers between 0 and 42 d of age. Plasma, liver, and muscle Se concentrations were improved by all Se sources at both d 21 and 42 compared with the negative control group. Between Se sources, minor differences were observed for plasma and liver results, whereas a significant dose effect was observed from 0.1 to 0.3 mg of Se/kg of feed (P < 0.05) for each source. Muscle Se concentrations were improved such as SO > SY > SS (P < 0.05). Moreover, the relative muscle Se enrichment comparison, using linear regression slope ratio, indicated an average of 1.48-fold (95% CI 1.38, 1.58) higher Se deposition in muscle for SO compared with SY. In the second experiment, excessive dietary doses of 5 mg of Se/kg of feed from SS and SO showed a lower deleterious effect of SO on BW and feed intake in comparison with standard Se doses (P < 0.05). Seleno amino acid measurements conducted on different tissues of animals fed SO at 0.5 mg/kg of feed showed that HMSeBA is fully converted into selenomethionine and selenocysteine. These results of both experiments demonstrate the higher relative bioavailability of SO compared with SS and SY as determined through tissue Se enrichment.

The methionine precursor DL-2-hydroxy-(4-methylthio)butanoic acid protects intestinal epithelial barrier function.

DL-2-hydroxy-(4-methylthio)butanoic acid (HMTBA) is a source of dietary methionine (Met) that is widely used in poultry nutrition. We have previously shown that HMTBA is preferentially diverted to the transsulfuration pathway, which gives antioxidant metabolites such as taurine and glutathione. Therefore, here we hypothesize that this Met source can protect epithelial barrier function in an in vitro model of intestinal inflammation of Caco-2 cells. The results show that HMTBA prevents the increase in paracellular permeability induced by H2O2 or tumour necrosis factor-α. This effect can be attributed to the increased production of taurine and reduced glutathione. Similar results were obtained for DL-Met, although the protective role of the amino acid was less pronounced than that of the hydroxy analogue. In conclusion, the diversion to the transsulfuration pathway means that this Met precursor is of greater value than previously thought, due to its capacity to improve intestinal homeostasis and the quality of poultry products destined for human consumption.

Comparative study of a new organic selenium source v. seleno-yeast and mineral selenium sources on muscle selenium enrichment and selenium digestibility in broiler chickens.

Two experiments were conducted on broiler chickens to compare the effect of a new organic Se source, 2-hydroxy-4-methylselenobutanoic acid (HMSeBA; SO), with two practical Se additives, sodium selenite (SS) and Se yeast (SY). The relative bioavailability of the different Se sources was compared on muscle (pectoralis major) total Se, selenomethionine (SeMet) and selenocysteine (SeCys) concentrations and apparent digestibility of total Se (ADSe). In the first experiment, from day (d) 0 to d21, Se sources were tested at different supplied levels and compared with an unsupplemented diet (NC). No significant effects were observed on growth performance during the experimental period. However, the different Se sources and levels improved muscle Se concentration compared with the NC, with a significant source effect in the following order: SS < SY < SO (P<0·05). Seleno-amino acids speciation results for NC, SY and SO at 0·3 mg Se/kg feed indicated that muscle Se was only present as SeMet or SeCys, showing a full conversion of Se by the bird. The second experiment (d0-d24) compared SS, SY or SO at 0·3 mg Se/kg feed. The ADSe measurements carried out between d20 and d23 were 24, 46 and 49% for SS, SY and SO, respectively, with significant differences between the organic and mineral Se sources (P<0·05). These results confirmed the higher bioavailability of organic Se sources compared with the mineral source and demonstrated a significantly better efficiency of HMSeBA compared with SY for muscle Se enrichment.

Effects of dietary protein content and 2-hydroxy-4-methylthiobutanoic acid or DL-methionine supplementation on performance and oxidative status of broiler chickens.

Besides its typical role as an amino acid in protein synthesis, methionine is an important intermediate in methylation reactions. In addition, it can also be converted to cysteine and hence plays a role in the defence against oxidative stress. The present study was conducted to investigate further the role of DL-methionine (DLM) and its hydroxy analogue, DL-2-hydroxy-4-methylthiobutanoic acid (DL-HMTBA), on zootechnical performance and oxidative status of broiler chickens. Male broiler chickens were reared on two diets differing in crude protein (CP) content (low-protein, 18·3 % v. high-protein, 23·2 % CP) and were supplemented either with 0·25 % DLM or 0·25 % DL-HMTBA. Reducing the dietary protein content resulted in an impaired body weight gain (P < 0·0001). However, supplementation of DL-HMTBA to the low-protein diet partially alleviated these negative effects (P = 0·0003). This latter phenomenon could be explained by the fact that chickens fed DL-HMTBA-supplemented diets displayed a better antioxidant status as reflected in lower lipid peroxidation probably as a consequence of their higher hepatic concentrations of total and reduced glutathione compared with their DLM counterparts. On the other hand, within the high protein levels, uric acid might be an important antioxidant to explain the lower lipid peroxidation of high-protein DL-HMTBA-supplemented chickens. Hepatic methionine sulfoxide reductase-A gene expression was not significantly affected by the dietary treatments. In conclusion, the present study indicates that there are interactions between dietary protein content and supplementation of methionine analogues with respect to broiler performance and antioxidant status, also suggesting a causal link between these traits.

An integrative in vitro approach to analyse digestion of wheat polysaccharides and the effect of enzyme supplementation.

The digestion of polysaccharides from the wheat cultivars Caphorn and Isengrain was investigated, and the efficiency of an enzyme preparation was tested using the TNO gastrointestinal model (TIM-1). The apparent digestibility (AD) of carbohydrates was determined based on the measurement of organic matter (OM), total monosaccharides, reducing ends (RE) and end products (EP: glucose, maltose and xylobiose). The AD of the OM from Caphorn and Isengrain measured using caecectomised cockerels did not differ from that measured using TIM-1: 72.0 (SD 2.6) v. 70.6 (SD 0.6) % for Caphorn (P = 0.580) and 73.0 (SD 2.3) v. 71.1 (SD 1.9) % for Isengrain (P = 0.252). After the 6 h TIM-1 digestion, 41.4-58.9 % of the OM, RE and EP were recovered from the jejunal compartment and 18.3-27.1 % from the ileal compartment, while ileal deliveries and digestive residues constituted the remainder. A commercial enzyme cocktail tested at 0.2 µl/g of wheat improved TIM-1 digestibility of Caphorn and Isengrain polysaccharides: 3.9 % (P = 0.0203) and 3.4 % (P = 0.0058) based on the OM; 9.7 % (P < 0.0001) and 3.1 % (P = 0.031) based on the total glucose; 47.2 % (P < 0.0001) and 14.2 % (P = 0.0004) based on the RE, respectively. The enzyme cocktail improved the release of the EP for Caphorn (3.8 %, P = 0.008) but not for Isengrain ( − 0.8 %, P = 0.561). The higher efficiency of the enzyme supplementation on the digestion of Caphorn polysaccharides compared with Isengrain seems to be linked to the higher soluble carbohydrate contents and/or less ramified arabinoxylan of Caphorn.

Intestinal cell conversion of DL-2-hydroxy-(4-methylthio)butanoic acid in vitro: dietary up-regulation by this methionine precursor.

DL-2-Hydroxy-(4-methylthio)butanoic acid (HMTBA) is a synthetic source of dietary methionine (Met) widely used in poultry nutrition. HMTBA is transported in the intestinal epithelium by the monocarboxylate transporter 1, after which its biological utilisation relies on its conversion to L-Met. This process involves stereospecific HMTBA oxidation to 2-keto-(4-methylthio)butanoic acid (KMB) and transamination to L-Met. In the present study, we examined HMTBA conversion to L-Met, further incorporation into cellular proteins and the regulation of both processes by HMTBA supplementation in differentiated intestinal Caco-2 cells. The results showed D- and L-HMTBA oxidation in the enterocytes, this process being up-regulated by HMTBA. The data also revealed that KMB transamination is not linked to a specific amino group donor. However, the branched-chain amino acid L-leucine is the preferred amino group donor. Furthermore, transamination was not affected by HMTBA availability. The incorporation of radioactivity from HMTBA into cellular proteins was not significantly different from that of L-Met and was not affected by HMTBA supplementation. In conclusion, the results reveal the capacity of Caco-2 cells to convert HMTBA to L-Met and the up-regulation of conversion by nutritional HMTBA supplementation, thus highlighting the contribution of the intestinal epithelium in the utilisation of HMTBA as a dietary source of Met.

Methionine deprivation regulates the S6K1 pathway and protein synthesis in avian QM7 myoblasts without activating the GCN2/eIF2 alpha cascade.

Amino acids modulate mRNA translation through the 70 kDa ribosomal protein S6 kinase (S6K1) and the general control nondepressible 2 protein kinase (GCN2)/eukaryotic initiation factor 2 alpha eIF2 alpha pathways. The aim of the present study was therefore to explore the signaling cascades potentially modulated by methionine availability in quail muscle QM7 myoblasts using media providing all other amino acids. Methionine deprivation caused a lower S6K1 phosphorylation compared with control (Ctl) cells. Supplying the methionine-deprived media with L- and DL-methionine isomers restored S6K1 phosphorylation to the levels observed in Ctl cells. Methionine also regulated downstream S6K1 targets (i.e. ribosomal protein S6 and eukaryotic elongation factor 2), modulated translation preinitiation complex (PIC) assembly, and stimulated protein synthesis. Replacing the lacking methionine with D-methionine or its hydroxyanalog [2-hydroxy-(4-methylthio) butanoic acid] did not restore S6K1 activation or protein synthesis. Conversely, the S6K1 pathway was activated by a methionine precursor, the ketoanalog of methionine. Methionine availability regulated the GCN2/eIF2 alpha pathway. However, our results indicate that methionine deprivation led to lower protein synthesis without activating eIF2 alpha phosphorylation, a process known to limit the formation of the 43S PIC. Using the amino acid alcohol methioninol did not decrease S6K1 phosphorylation or activity and did not alter the regulation of protein synthesis by methionine. These findings suggest that methionine exerts an effect on S6K1 signaling and protein synthesis in avian QM7 myoblasts through a mechanism partly independent of the global regulation via tRNA charging.

Properties of selected hemicellulases of a multi-enzymatic system from Penicillium funiculosum.

A multi-enzymatic system from Penicillium funiculosum displayed alpha-L-arabinofuranosidase, endo-1,4-beta-D-xylanase, beta-D-xylosidase and endo-1,3-1,4-beta-D-glucanase activities at high levels over a wide acidic pH range of 2.0 to 5.5. Moreover, the pH stability was particularly extended over the wide range of pH of 2.0 to 8.0 with endo-1,3-1,4-beta-D-glucanase and endo-1,4-beta-D-xylanase; however, alpha-L-arabinofuranosidase and beta-D-xylosidase exhibited higher stability in the pH range of 2.0 to 5.5. The results indicate that the optimal temperature of alpha-L-arabinofuranosidase (65 degrees C) and beta-D-xylosidase (70 degrees C) as well as their thermal stability were higher than those of endo-1,3-1,4-beta-D-glucanase (60 degrees C) and endo-1,4-beta-D-xylanase (50 degrees C). Although V(maxapp) of beta-D-xylosidase and endo-1,4-beta-D-xylanase was higher than that of alpha-L-arabinofuranosidase and endo-1,3-1,4-beta-D-glucanase, respectively, their catalytic efficiency was lower. High levels of ferulolyl esterase, alpha-D-galactosidase, beta-D-mannosidase and endo-1,4-beta-D-mannanase activities were also detected in the multi-enzymatic system. The overall features of the multi-enzymatic system from P. funiculosum reveal its potential for degrading and modifying plant cell walls from a variety of food and feedstuffs.

Effect of pH on L- and D-methionine uptake across the apical membrane of Caco-2 cells.

The transport systems involved in intestinal methionine (Met) absorption are described as Na(+)-dependent and Na(+)-independent mechanisms. However, since recent studies have suggested the importance of the H(+) gradient as a driving force for intestinal nutrient absorption, the aim of the present work was to test whether Met transport across the apical membrane of Caco-2 cells is affected by extracellular pH. The results show that l- and d-Met uptake was increased by lowering extracellular pH from 7.4 to 5.5, in both the presence and absence of Na(+). Cis-inhibition experiments revealed that inhibition of l-Met transport by 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid (BCH) or l-lysine (l-Lys) was higher at a pH of 5.5. Moreover, the BCH-insensitive component was not affected by pH, whereas the l-Lys-insensitive component was increased by lowering extracellular pH, thus suggesting the participation of system L. The contribution of another mechanism, sensitive to both BCH and l-Lys, was also considered. The inhibition obtained with taurine (Tau) was also higher at a pH of 5.5, thus suggesting the involvement of system B(0,+) on pH-stimulated component. As for d-Met uptake, the results showed higher inhibition with l-Lys and Tau at a pH of 5.5 and no effect on the l-Lys- or Tau-insensitive component. In conclusion, Met transport across the apical membrane of Caco-2 cells is increased by low extracellular pH as the result of the stimulation of two transport systems functionally identified with systems L and B(0,+) for l-Met and with system B(0,+) for d-Met.

Characterization of selected cellulolytic activities of multi-enzymatic complex system from Penicillium funiculosum.

The presence of endo-1,4-beta-D-glucanase, cellobiohydrolase, and beta-glucosidase activities in a multi-enzymatic complex system from Penicillium funiculosum was investigated. The interesting feature of these enzymes is their synergistic action for the hydrolysis of the native cellulose into glucose units. Both endo-1,4-beta-D-glucanase and cellobiohydrolase showed broader pH activity profiles, with pH optima of 4.0 and 4.0-5.0, respectively. However, beta-glucosidase activity showed a narrow pH-activity profile, with an optimum pH of 4.5. The different cellulolytic activities were stable in the acidic pH range of 2.5-6.0 and showed a similar optimal temperature of 60 degrees C. Although beta-glucosidase has shown a close catalytic efficiency as that of endo-1,4-beta-D-glucanase, its thermal stability was lower. However, the thermal stability profile of cellobiohydrolase was close to that of endo-1,4-beta-D-glucanase. The results also revealed the presence of high levels of endo-1,3-1,4-beta-D-glucanase, endo-1,3-beta- d-glucanase, and pectinase activities in the multi-enzymatic cellulolytic complex system. Moreover, the investigated multi-enzymatic complex system was effective in degrading the nonstarch polysaccharides of soybean meal.

Mechanisms through which sulfur amino acids control protein metabolism and oxidative status.

Amino acids regulate protein synthesis and breakdown (i.e., protein turnover) and consequently protein deposition, which corresponds to the balance between the two processes. Elucidating the mechanisms involved in such regulation is important from fundamental and applied points of view since it can provide a basis to optimize amino acid requirements and to control protein mass, body composition and so forth. Amino acids, which have long been considered simply as precursors of protein synthesis, are now recognized to exert other significant influences; that is, they are precursors of essential molecules, act as mediators or signal molecules and affect numerous functions. For example, amino acids act as mediators of metabolic pathways in the same manner as certain hormones. Thus, they modulate the activity of intracellular protein kinases involved in the regulation of metabolic pathways such as mRNA translation. We provide here an overview of the roles of amino acids as regulators of protein metabolism, by focusing particularly on sulfur amino acids. The potential importance of methionine as a "nutrient signal" is discussed in the light of recent findings. Emphasis is also placed on mechanisms controlling oxidative status since sulfur amino acids are involved in the synthesis of intracellular antioxidants (glutathione, taurine etc.) and in the methionine sulfoxide reductase antioxidant system.