Prediction of the amino acid digestibility of legume seeds in growing pigs: a meta-analysis approach

On pig farms, a high proportion of the cost of production comes from feed costs. However, the use of alternative ingredients such as legume seeds may help to reduce this cost. In fact, legume seeds are an important source of essential amino acids (EAA) and can therefore be an alternative to oilseed meals. However, the accurate use of these legume seeds requires a precise knowledge of the standardized ileal digestibility (SID) of EAA, which may vary depending on its botanical variety. A meta-analysis was performed on a database compiling data from 41 studies published between 1981 and 2013 and 178 dietary treatments. Models of prediction of the SID of EAA as well as the dietary concentration of digestible standardized EAA (dEAA) were obtained, based on the chemical composition of ingredients reported in the publications. The effect of the type of legume seeds (faba bean, lupin, pea and soya bean), surgical procedures (T-cannula, re-entrant cannulas, post valve T-cannulas and ileo-rectal anastomosis), and BW of pigs (BW⩽25 kg BW>25 kg) were also tested in each model. Results showed that dietary CP and crude fibre (CF) were, respectively, the best predictors of each EAA SID for faba bean, lupin and pea (R²=0.42 to 0.89) and soya bean (R²=0.32 to 0.77). For the dEAA content, the best prediction models included dietary CP and ADF for faba bean, lupin and pea and soya bean, respectively, with R² ranging from 0.66 to 0.98. Models developed in this study allow predicting the digestibility of EAA in these alternatives feedstuffs.     

Standardized ileal digestibility of amino acids in eight genotypes of barley fed to growing pigs

To determine chemical composition, physical characteristics and standardized ileal digestibility (SID) of CP and amino acids (AA) in eight current hulled barley genotypes, an experiment with growing pigs has been conducted. These genotypes included Yool, Campanile, Lomerit, Travira, Anisette, Canberra, Metaxa and Fridericus. Growing barrows with an average initial BW of 30±2 kg were surgically fitted with simple T-cannulas at the distal ileum, and allotted to an 8×9 Youden square design with eight periods of 6 days each and nine pigs. Barley was the sole dietary source of CP and AA. On average, the eight genotypes contained on as-fed basis 10.7% CP, 15.2% total non-starch polysaccharides (NSP), 17.1% NDF and 4.1% β-glucan, and had a mean test weight (TW) of 72.2 kg/hl. The SID of CP in the barley genotypes varied from 69% to 74%, and was greater (P<0.01) for genotypes Travira, Anisette and Metaxa compared to Yool and Campanile. Standardized ileal digestibility of Lys, Met and Trp (P<0.05) but not of Thr differed between genotypes. Moreover, barley genotypes differed in their standardized ileal digestible content (cSID) of CP and AA. Furthermore, SID and cSID of CP and most AA linearly decreased (P<0.05) with increasing NDF and total sugar content. Standardized ileal digestibility of CP and some AA and cSID of CP and most AA decreased linearly with increasing TW (P<0.05). Additionally, SID and cSID of CP and AA of most barley genotypes were lower when compared to tabulated values. In conclusion, a comprehensive database on chemical composition and SID of CP and AA in eight current barley genotypes has been made available. However, as present SID values are lower compared to feed tables, adjustments are required to minimize the risk of overestimating the actual protein value of barley for pigs.     

Nutrient and energy balance,and amino acid digestibility in weaned piglets fed wheat bran and an exogenous enzyme combination

The digestive system of the weaned piglets can be affected by the type of ingredients present in the diet, and a high fibre content in diets can limit the use of other nutrients and energy. The study was conducted to determine the effects of multicarbohydrase (MC) and phytase (Phy) supplementation on the nutritive value of wheat bran (WB) in weaned piglets. Multicarbohydrase preparation had 700 U α-galactosidase, 2200 U galactomannanase, 3000 U xylanase and 22 000 U β-glucanase per kilogram of diet, and Phy had 500 phytase units – FTU/kg of diet. Twenty-five weaned piglets (6.1 ± 0.63 kg) at 21 days old were fed five diets in a completely randomised experimental design with a 2 × 2 + 1 (0 and 200 mg/kg MC; 0 and 50 mg/kg Phy; and basal diet – BD) factorial arrangement used to determine treatment effects. An additional group of piglets was fed a corn-basal diet during apparent digestibility of nutrients, and fed a 5% casein-corn starch basal diet during apparent and standardised ileal digestibility (SID) of amino acid evaluations. Piglets were individually caged until 38 days old, when Ileal digesta was collected at slaughter. Test diets were made by mixing the basal diets and WB 7 : 3 (w/w), with or without MC, Phy or the combination. There was an interaction trend (P = 0.07) between MC and Phy in the balance of ash, digestible energy (DE) and metabolisable energy (ME). Effects of MC (P < 0.01) on DM, N retention, DE and ME, as well as an effect of Phy (P < 0.05) on ash, DE and ME and a trend in protein digestibility (P = 0.07) also was observed. The enzyme combination showed effect (P < 0.05) on SID of Lys, Pro and Ser; as a trend (P < 0.07) on His, Thr and Val. Isolated, MC also suggested improving (P < 0.07) on SID of His, Lys, Ala (P < 0.05), while Phy improved (P < 0.06) SID of Leu, Lys, Met (P < 0.01), Thr, Val, Ala (P < 0.01), Pro and Ser (P < 0.05). The MC carbohydrate complex was characterised as a viable alternative to increase the apparent nutrients digestibility and SID of amino acids when WB was used in the diet of young pigs and, when combined with Phy, suggested an additive effect on the apparent use of energy.     

The primary structure of the β-subunit of protocatechuate 3,4-dioxygenase from Pseudomonas aeruginosa

The complete amino acid sequence of the β-subunit of protocatechuate 3,4-dioxygenase was determined. The β-subunit contained four methionine residues. Thus, five peptides were obtained after cleavage of the carboxymethylated β-subunit with cyanogen bromide, and were isolated on Sephadex G-75 column chromatography. The amino acid sequences of the cyanogen bromide peptides were established by characterization of the peptides obtained after digestion with trypsin, chymotrypsin, thermolysin, or Staphylococcus aureus protease. The major sequencing techniques used were automated and manual Edman degradations. The five cyanogen bromide peptides were aligned by means of the amino acid sequences of the peptides containing methionine purified from the tryptic hydrolysate of the carboxymethylated β-subunit. The amino acid sequence of all the 238 residues was as follows: ProAlaGlnAspAsnSerArgPheValIleArgAsp ArgAsnTrpHis ProLysAlaLeuThrPro-Asp — TyrLysThrSerIleAlaArg SerProArgGlnAla LeuValSerIleProGlnSer — IleSerGluThrThrGly ProAsnPheSerHisLeu GlyPheGlyAlaHisAsp-His — AspLeuLeuLeuAsnPheAsn AsnGlyGlyLeu ProIleGlyGluArgIle-Ile — ValAlaGlyArgValValAsp GlnTyrGlyLysPro ValProAsnThrLeuValGluMet — TrpGlnAlaAsnAla GlyGlyArgTyrArg HisLysAsnAspArgTyrLeuAlaPro — LeuAspProAsn PheGlyGlyValGly ArgCysLeuThrAspSerAspGlyTyrTyr — SerPheArg ThrIleLysProGlyPro TyrProTrpArgAsnGlyProAsnAsp — TrpArgProAla HisIleHisPheGlyIle SerGlyProSerIleAlaThr-Lys — LeuIleThrGlnLeuTyr PheGluGlyAspPro LeuIleProMetCysProIleVal — LysSerIleAlaAsn ProGluAlaValGlnGln LeuIleAlaLysLeuAspMetAsnAsn — AlaAsnProMet AsnCysLeuAlaTyr ArgPheAspIleValLeuArgGlyGlnArgLysThrHis PheGluAsnCys. The sequence published earlier in summary form (Iwaki et al., 1979, J. Biochem.86, 1159–1162) contained a few errors which are pointed out in this paper.     

Sequences of tryptic peptides containing the five cysteinyl residues of ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum

Tryptic peptides which account for all five cysteinyl residues in ribulosebisphosphate carboxylase/oxygenase from Rhodospirillum rubrum have been purified and sequenced. Collectively, these peptides contain 94 of the approximately 500 amino acid residues per molecule of subunit. Due to one incomplete cleavage at a site for trypsin and two incomplete chymotryptic-like cleavages, eight major radioactive peptides (rather than five as predicted) were recovered from tryptic digests of the enzyme that had been carboxymethylated with [³H]iodoacetate. The established sequences are: GlyTyrThrAlaPheValHisCys¹Lys TyrValAspLeuAlaLeuLysGluGluAspLeuIleAla GlyGlyGluHisValLeuCys¹AlaTyr AlaGlyTyrGlyTyrValAlaThrAlaAlaHisPheAla AlaGluSerSerThrGlyThrAspValGluValCys¹ ThrThrAsxAsxPheThrArg AlaCys¹ThrProIleIleSerGlyGlyMetAsnAla LeuArg ProPheAlaGluAlaCys¹HisAlaPheTrpLeuGly GlyAsnPheIleLys In these peptides, radioactive carboxymethylcysteinyl residues are denoted with asterisks and the sites of incomplete cleavage with vertical wavy lines. None of the peptides appear homologous with either of two cysteinyl-containing, active-site peptides previously isolated from spinach ribulosebisphosphate carboxylase/oxygenase.     

Dose–response evaluation of the standardized ileal digestible tryptophan : lysine ratio to maximize growth performance of growing-finishing gilts under commercial conditions

Environmental regulations as well as economic incentives have resulted in greater use of synthetic amino acids in swine diets. Tryptophan is typically the second limiting amino acid in corn-soybean meal-based diets. However, using corn-based co-products emphasizes the need to evaluate the pig’s response to increasing Trp concentrations. Therefore, the objective of these studies was to evaluate the dose–response to increasing standardized ileal digestible (SID) Trp : Lys on growth performance of growing-finishing gilts housed under large-scale commercial conditions. Dietary treatments consisted of SID Trp : Lys of 14.5%, 16.5%, 18.0%, 19.5%, 21.0%, 22.5% and 24.5%. The study was conducted in four experiments of 21 days of duration each, and used corn-soybean meal-based diets with 30% distillers dried grains with solubles. A total of 1166, 1099, 1132 and 975 gilts (PIC 337×1050, initially 29.9±2.0 kg, 55.5±4.8 kg, 71.2±3.4 kg and 106.2±3.1 kg BW, mean±SD) were used. Within each experiment, pens of gilts were blocked by BW and assigned to one of the seven dietary treatments and six pens per treatment with 20 to 28 gilts/pen. First, generalized linear mixed models were fit to data from each experiment to characterize performance. Next, data were modeled across experiments and fit competing dose–response linear and non-linear models and estimate SID Trp : Lys break points or maximums for performance. Competing models included broken-line linear (BLL), broken-line quadratic and quadratic polynomial (QP). For average daily gain (ADG), increasing the SID Trp : Lys increased growth rate in a quadratic manner (P<0.02) in all experiments except for Exp 2, for which the increase was linear (P<0.001). Increasing SID Trp : Lys increased (P<0.05) feed efficiency (G : F) quadratically in Exp 1, 3 and 4. For, ADG the QP was the best fitting dose–response model and the estimated maximum mean ADG was obtained at a 23.5% (95% confidence interval (CI): [22.7, 24.3%]) SID Trp : Lys. For maximum G : F, the BLL dose–response models had the best fit and estimated the SID Trp : Lys minimum to maximize G : F at 16.9 (95% CI: [16.0, 17.8%]). Thus, the estimated SID Trp : Lys for 30 to 125 kg gilts ranged from a minimum of 16.9% for maximum G : F to 23.5% for maximum ADG.     

Requirement of standardized ileal digestible valine to lysine ratio for 8- to 14-kg pigs

The objective was to define the Val requirement for weaned piglets in the context of reducing the dietary protein content. A dose–response experiment was conducted to estimate the standardized ileal digestible (SID) Val to Lys ratio required to support the optimum growth of post-weaned piglets. In this study, 96 pigs weighing 8 kg were allotted to one of six dietary treatments (16 pigs for each dietary treatment) and were housed individually. Diets were formulated to provide 0.58, 0.62, 0.66, 0.70, 0.74 and 0.78 SID Val : Lys by adding graded levels of crystallinel-Val to the 0.58 SID Val : Lys diet. Lysine was sub-limiting and supplied 90% of the recommendation (10.95 g SID Lys/kg equal to 11.8 g/kg total Lys). Average daily feed intake (ADFI), average daily gain (ADG) and gain to feed ratio (G : F) were determined during a 14-day period ofad libitum feeding. Blood and urine samples were taken at the end of each week (day 7 and 14 of the experiment) 3 h after feeding the experimental diets. The maximum ADFI and ADG were obtained in pigs fed the 0.78 SID Val : Lys diet; it was not different from the results of pigs fed 0.70 SID Val : Lys diet. The highest G : F was obtained in pigs fed 0.70 SID Val : Lys. The plasma concentration of Val increased linearly (P<0.001) as the dietary SID Val : Lys increased. The increasing dietary Val : Lys also resulted in a linear increase in Cys (P<0.001) and a quadratic increase in Arg (P=0.003), Lys (P=0.05) and Phe (P=0.009). The plasma Gly showed a quadratic decrease (P=0.05) as the dietary Val : Lys increased. Neither plasma nor urinary urea to creatinine ratio was affected by treatment. The minimum SID Val : Lys required to maximize ADFI, ADG and G : F was estimated at 0.67 SID Val : Lys by a broken-line model, and at 0.71 SID Val : Lys by a curvilinear plateau model. The Val deficiency caused a reduction in ADFI, and Val supplementation above the requirement did not impair animal performance. In conclusion, 0.70 SID Val : Lys is suggested as the Val requirement for 8 to 14 kg individually housed pigs.     

Impact of barley form on equine total tract fibre digestibility and colonic microbiota

This study aimed at assessing the impact of four barley forms on total tract apparent digestibility of dietary fibre in horses fed a large amount of starch in the morning meal (0.27% BW). Processed barley forms had a greater pre-caecal starch digestibility than the whole form. Based on this result, we hypothesised that using barley-processing methods would limit the potential dumping of undegraded starch in the hindgut of horses and, consequently, the potential negative effect on fibre degradation in the hindgut. In a 4×4 latin square design, four mature geldings fitted with a right ventral colon-fistula were fed a meadow hay : concentrate (62 : 38; dry matter (DM) basis) diet at 1.7% BW. The concentrate was made of 80% barley distributed either as whole grain or as processed forms: 2.5 mm ground, pelleted or steam-flaked. For each period, total tract apparent digestibilities of DM, NDF and ADF were determined over 3 consecutive days by total faecal collection, whereas pH, volatile fatty acids (VFA) concentrations and cultural functional bacteria counts (total anaerobic, cellulolytic bacteria, lactic acid producers, amylolytic bacteria and lactic acid utilisers) in colonic content were evaluated on 1 day 4 h after the morning meal. Total tract apparent digestibility of DM and dietary fibre was influenced (P<0.05) by barley form. Diets including thermo-mechanically treated barley forms led to a higher (P<0.05) total tract apparent digestibility of NDF than those constituted of ground barley and also led to a greater (P<0.05) total tract apparent digestibility of ADF than those made of whole or ground barley forms. However, no significant difference was observed in colonic pH, VFA concentrations and cultural bacteria concentrations. Owing to a high starch supply in the morning meal, the concentration of the functional bacteria in the colonic content averaged 7.8 log CFU/ml, 5.9 NPM/ml, 6.9 and 7.3 CFU/ml for total anaerobic, cellulolytic, amylolytic and lactic acid-utilising bacteria, respectively. Consequently, providing horses with pelleted or steam-flaked instead of ground barley forms may limit the negative impact of starch on fibre digestibility in horses fed a high level of starch in the morning meal (0.27% BW). Moreover, the fibre-to-starch ratio fed in this experiment did not cause any digestive upset.     

NH2-terminal sequences of DNA-, heparin-, and gelatin-binding tryptic fragments from human plasma fibronectin

NH₂-terminal sequence analysis was performed on subregions of human plasma fibronectin including 24,000-dalton (24K) DNA-binding, 29,000-dalton (29K) gelatin-binding, and 18,000-dalton (18K) heparin-binding tryptic fragments. These fragments were obtained from fibronectin after extensive trypsin digestion followed by sequential affinity purification on gelatin-Sepharose, heparin-agarose, and DNA-cellulose columns. The gelatin-binding fragment was further purified by gel filtration on Sephadex G-100, and the DNA-binding and heparin-binding fragments were further purified by high-performance liquid chromatography. The 29K fragment had the following NH₂-terminal sequence: AlaAlaValTyrGlnProGlnProHisProGlnProPro (Pro)TyrGlyHis HisValThrAsp(His)(Thr)ValValTyrGly(Ser) ?(Ser)?-Lys. The NH₂-terminal sequence of a 50K, gelatin-binding, subtilisin fragment by L. I. Gold, A. Garcia-Pardo, B. Prangione, E. C. Franklin, and E. Pearlstein (1979, Proc. Nat. Acad. Sci. USA76, 4803–4807) is identical to positions 3–19 (with the exception of some ambiguity at position 14) of the 29K fragment. These data strongly suggest that the 29K tryptic fragment is included in the 50K subtilisin fragment, and that subtilisin cleaves fibronectin between the Ala²Val³ residues of the 29K tryptic fragment. The 18K heparin-binding fragment had the following NH₂-terminal sequence: (Glu)AlaProGlnProHisCysIleSerLysTyrIle LeuTyrTrpAspProLysAsnSerValGly?(Pro) LysGluAla?(Val)(Pro). The 29K gelatin-binding and 18K heparin-binding fragments have proline-rich NH₂-terminal sequences suggesting that they may have arisen from protease-sensitive, random coil regions of fibronectin corresponding to interdomain regions preceding macromolecular-binding domains. Both of these fragments contain the identical sequence ProGlnProHis, a sequence which may be repeated in other interdomain regions of fibronectin. The 24K DNA-binding fragment has the following NH₂-terminal sequence: SerAspThrValProSerProCysAspLeuGlnPhe ValGluValThrAspVal LysValThrIleMetTrpThrProProGluSerAla ValThrGlyTyrArgVal AspValCysProValAsnLeuProGlyGluHisGly Gln(Cys)LeuProIleSer. The sequence of positions 9–22 are homologous to positions 15–28 of the α chain of DNA-dependent RNA polymerase from Escherichia coli. The homology observed suggests that this stretch of amino acids may be a DNA-binding site.     

Determination of standardized ileal crude protein and amino acid digestibilities in protein supplements for piglets

The objective of the study was to determine standardized ileal digestibilities (SID) of crude protein (CP) and amino acids (AA) in six ingredients commonly used in weanling pigs diets. The ingredients consisted of extruded soybeans (SBe), high-protein soybean meal (SBMhp), soy protein concentrate (SPC), hydrolyzed wheat gluten (WGh), conventional fish meal 1 (FM1) and extracted fish meal 2 (FM2e). Each assay feed ingredient was added to a purified cornstarch-based basal diet supplemented with casein and crystalline AA, and SID of CP and AA of assay ingredients were estimated using the difference method. A total of 12 3-week-old barrows were surgically fitted with T-cannulas at the distal ileum. The piglets were allocated to four simultaneous 3 × 3 Latin-square designs with three animals and three periods each, resulting in six observations per assay diet. After 3 to 5 days recovery from surgery, each assay diet was fed at a level of 30 g/kg body weight. Each experimental period consisted of 5 days for adaptation to the assay diets and 2 days for ileal digesta collection. The SID of CP, Lys, Met, Thr and Trp were 73%, 78%, 76%, 66% and 71% in SBe; 80%, 84%, 87%, 75% and 81% in SBMhp; 86%, 89%, 89%, 78% and 83% in SPC; 87%, 60%, 88%, 76% and 79% in WGh; 87%, 92%, 94%, 86% and 86% in FM1; and 79%, 86%, 89%, 80% and 74% in FM2e, respectively. The greatest SID of indispensable AA were observed for FM1 and SPC (SID 85% for most indispensable AA), followed by WGh, FM2e and SBMhp (SID 80% for most indispensable AA) and the smallest SID were obtained for SBe (SID < 80% for most indispensable AA). The SID of CP and indispensable AA in both types of fish meal, SBMhp and WGh were similar to SPC (P = 0.063 to 0.855), except for Arg (P = 0.010) and Lys (P = 0.001) in WGh, and Phe (P = 0.044) and Trp (P = 0.037) in FM2e. The SID of CP and indispensable AA were smaller in SBe compared with SPC (P < 0.001 to P = 0.017), except for Lys (P = 0.136). The SID of CP and indispensable AA were consistently smaller in FM2e compared with FM1 with significant differences for CP (P = 0.035), Phe (P = 0.028) and Trp (P = 0.008). Digestibility values measured in the present study can be used to formulate diets for piglets based on standardized ileal digestible contents of CP and AA.     

Effects of orally administrated amino acids on myofibrillar proteolysis in chicks

We examined the effects of orally administrated amino acids on myfibrillar proteolysis in food-deprived chicks. Plasma N(tau)-methylhistidine concentration, as an index of myofibrillar proteolysis, was decreased by the administration of Glu, Gly, Ala, Leu, Ile, Ser, Thr, Met, Trp, Asn, Gln, Pro, Lys and Arg but not by Asp, Val, Phe, Tyr or His to chicks. Orally administrated Cys was fatal to chicks. These results indicate that oral Glu, Gly, Ala, Leu, Ile, Ser, Thr, Met, Trp, Asn, Gln, Pro, Lys and Arg administration suppressed myofibrillar proteolysis in chicks.     

Effect of partial dehulling of two- and six-row barley varieties on precaecal digestibility of amino acids in pigs

Five barrows (German Landrace; initial BW 58 kg, final BW 80 kg) fitted with an ileo-rectal anastomosis were used to determine the effect of partial dehulling and addition of barley hulls of two- and six-row barley varieties on the precaecal digestibility (pD) of CP and amino acids. The following diets were provided according to a standardized diet formulation and tested in seven consecutive periods (repeated group-period design): two-row barley (TRB) + casein (C), dehulled TRB + C, TRB + C + 10% hulls, six-row barley (SRB) + C, dehulled SRB + C, SRB + C + 1% hulls, and wheat starch + C. The diets were supplied at daily rates of 79-86 g DMI x kg BW(-0.75) in barley containing diets and at 49 g DMI x kg BW(-0.75) in the casein diet. The digestibility of amino acids in barley varieties was determined by the difference method (casein as basal diet) using quantitative digesta collection. In both varieties of barley the pD of CP and amino acids did not differ. The pD of CP was unchanged in regard to the treatments in both barley varieties. Due to dehulling in TRB the pD was improved significantly for most indispensable amino acids and in SRB for Met and Cys. Addition of 10% hulls to TRB led to equivalent pD of Arg, His, Leu, Tyr, and Trp compared to TRB, but the pD of Lys, Phe, Thr and Val was significantly decreased below the levels of TRB. Addition of even 1% hulls to SRB impaired the pD of Lys below the level in SRB. In conclusion, addition of barley hulls to pig diets impairs amino acids absorption in the small intestine. The pD values, measured under standardized experimental conditions (without a correction using basal endogenous amino acids), are similar to the values of true digestibility published by NRC (1998).     

Mutations in Intron 1 and Intron 22 Inversion Negative Haemophilia A Patients from Western India

Despite increased awareness and diagnostic facilities, 70–80% of the haemophilia A (HA) patients still remain undiagnosed in India. Very little data is available on prevalent mutations in HA from this country. We report fifty mutations in seventy one Indian HA patients, of which twenty were novel. Ten novel missense mutations [p.Leu11Pro (p.Leu-8Pro), p.Tyr155Ser (p.Tyr136Ser), p.Ile405Thr (p.Ile386Thr), p.Gly582Val (p.Gly563Val) p.Thr696Ile (p.Thr677Ile), p.Tyr737Cys (p.Tyr718Cys), p.Pro1999Arg (p.Pro1980Arg), p.Ser2082Thr (p.Ser2063Thr), p.Leu2197Trp (p.Leu2178Trp), p.Asp2317Glu (p.Asp2298Glu)] two nonsense [p.Lys396* (p.Lys377*), p.Ser2205* (p.Ser2186*)], one insertion [p.Glu1268_Asp1269ins (p.Glu1249_Asp1250)] and seven deletions [p.Leu882del (p.Leu863del), p.Met701del (p.Met682del), p.Leu1223del (p.Leu1204del), p.Trp1961_Tyr1962del (p.Trp1942_Tyr1943del) p.Glu1988del (p.Glu1969del), p.His1841del (p.His1822del), p.Ser2205del (p.Ser2186del)] were identified. Double mutations (p.Asp2317Glu; p.Thr696Ile) were observed in a moderate HA case. Mutations [p. Arg612Cys (p.Arg593Cys), p.Arg2326Gln (p.Arg2307Gln)] known to be predisposing to inhibitors to factor VIII (FVIII) were identified in two patients. 4.6% of the cases were found to be cross reacting material positive (CRM+ve). A wide heterogeneity in the nature of mutations was seen in the present study which has been successfully used for carrier detection and antenatal diagnosis in 10 families affected with severe to moderate HA.     

Amino acid utilization and body composition of growing pigs fed processed soybean meal or rapeseed meal with or without amino acid supplementation

Feed ingredients used in swine diets are often processed to improve nutritional value. However, (over-)processing may result in chemical reactions with amino acids (AAs) that decrease their ileal digestibility. This study aimed to determine effects of (over-)processing of soybean meal (SBM) and rapeseed meal (RSM) on post-absorptive utilization of ileal digestible AAs for retention and on body AA composition of growing pigs. Soybean meal and RSM were processed by secondary toasting in the presence of lignosulfonate to obtain processed soybean meal (pSBM) and processed rapeseed meal (pRSM). Four diets contained SBM, pSBM, RSM or pRSM as sole protein source. Two additional diets contained pSBM or pRSM and were supplemented with crystalline AA to similar standardized ileal digestible (SID) AA level as the SBM or RSM diet. These diets were used to verify that processing affected AA retention by affecting ileal AA digestibility rather than post-absorptive AA utilization. The SID AA levels of the protein sources were determined in a previous study. In total, 59 pigs were used (initial BW of 15.6±0.7 kg) of which five were used to determine initial body composition at the start of the experiment. In total, 54 pigs were fed one of six experimental diets and were slaughtered at a BW of 40 kg. The organ fraction (i.e. empty organs plus blood) and carcass were analyzed separately for N and AA content. Post-absorptive AA utilization was calculated from AA retention and SID AA intake. An interaction between diet type, comprising effects of processing and supplementing crystalline AA, and protein source was observed for CP content in the organ fraction, carcass and empty body and for nutrient retention. Processing reduced CP content and nutrient retention more for SBM than for RSM. Moreover, processing reduced (P<0.001) the lysine content in the organ fraction for both protein sources. Supplementing crystalline AA ameliorated the effect of processing on these variables. Thus, the data indicated that processing affected retention by reducing digestibility. Correcting AA retention for SID AA intake was, therefore, expected to result in similar post-absorptive AA utilization which was observed for the RSM diets. However, post-absorptive AA utilization was lower for the pSBM diet than for the SBM diet which might be related to an imbalanced post-absorptive AA supply. In conclusion, processing negatively affected nutrient retention for both protein sources and post-absorptive utilization of SID AA for retention for SBM. Effects of processing were compensated by supplementing crystalline AA.     

Evaluation through literature data on standardized ileal digestibility and basal ileal endogenous loss of amino acids associated with barley in pigs

This meta-analysis aimed to estimate the standardized ileal digestibility (SID) and the basal ileal endogenous amino acid losses (IAA_end) in barley for growing pigs. In total, 38 different barley treatments published in 26 peer-reviewed papers were used for the meta-analysis containing information on dietary composition including amino acid (AA) contents of the assay diets, and (or) barley samples, as well as apparent ileal digestibility (AID) of AA in barley. The SID of AA was determined by either correcting AID of AA for their IAA_end or by regression analysis between the apparent ileal digestible and total dietary AA contents. The SID values obtained by correcting the AID values for their IAA_end amounted to 70%, 77%, 74% and 63% for Lys, Met, Thr and Trp, whereas those based on regression analysis method were 82%, 82%, 69% and 55%, respectively. Estimates of basal ileal endogenous loss of CP in ileal digesta varied considerably and averaged 11.84 g/kg dry matter intake (DMI), whereas IAA_end for indispensable AA ranged from 0.05 g/kg DMI for Trp to 1.90 g/kg DMI for Leu. In most cases, these estimates were considerably higher than previously reported values for IAA_end. The results of the present regression analysis indicate for most AA higher SID values compared with SID of most AA that were obtained by correcting AID values for IAA_end. In view of the observed high variations in IAA_end and the low CP content of the barley samples, estimating SID of AA based on literature data by means of the regression method may improve accuracy of SID coefficients for barley. In contrast, transformation of AID values into their corresponding SID values by using a constant correction factor for IAA_end adds an additional source of error, thereby reducing the precision in estimating SID of AA.     

Ileal and total tract nutrient digestibility in blue foxes (Alopex lagopus) fed extruded diets containing different protein sources

This study compared apparent ileal and total tract digestibility of macronutrients and amino acids in blue foxes (Alopex lagopus) fed dry extruded diets. The control diet contained fish meal as the main protein source, while in the other diets 50% of the fish meal protein was replaced by not de-hulled solvent-extracted soybean meal, meat meal or bacterial protein meal. Ileal digestibility was obtained with animals modified with ileorectal anastomosis (IRA method). There were no significant differences in the average digestibility of CP, Lys, Trp, Val, Ala or Glu between IRA modified and intact animals. Average ileal digestibilities of His, Thr, Asp, Cys, Gly, Hyp, Pro, Ser, starch and total carbohydrates (CHO) were significantly lower compared with total tract values. Average ileal digestibility of Arg, Ile, Leu, Met, Phe, Tyr and crude fat was significantly higher than total tract digestibility. Digestibility measured in IRA modified animals showed less variability compared with values from intact animals. There were significant differences among diets in ileal as well as total tract digestibility of orude protein, most amino acids, starch and CHO, and in ileal digestibility of crude fat. Ileal crude protein digestibility of the diets ranged from 81.0-86.4% and total tract digestibility from 82.5-86.4%. Ileal amino acid digestibilities ranged from 70.1 (Asp) to 93.3% (Arg) and total tract values ranged from 72.8 (Cys) to 92.2% (Arg). Both ileal and total tract digestibilities of crude protein and most amino acids were significantly lower for diets containing meat meal or bacterial protein meal compared with the control fish meal-based diet and the diet with soybean meal. Although ileal digestibility may be more accurate than total tract digestibility in estimating the protein and amino acid supply in blue foxes, total tract digestibility may be acceptable because of numerically small differences between ileal and total tract digestibilities for protein and most amino acids. It was further concluded that ileal digestibilities of starch and CHO in the blue fox are lower than corresponding total tract digestibilities.     

Improving the estimation of amino acid requirements to maximize nitrogen retention in precision feeding for growing-finishing pigs

Precision feeding requires a mathematical model to estimate standardized ileal digestible (SID) lysine (Lys) requirements (SIDLysR) in real time. However, this type of model requires constant calibration updates. The objective of this study was to review the calibration of the model used to estimate the real-time Lys requirements of individual growing-finishing pigs. A digestibility trial (n = 10) was conducted to evaluate amino acids digestibility during the growing and finishing phases. Additionally, 120 pigs were used in two 28-day growth experiments conducted as completely randomized design with growing (25 ± 2.1 kg BW, n = 60; 10 pigs per treatment) or finishing barrows (68.1 ± 6 kg BW, n = 60; 10 pigs per treatment). In each experiment, the pigs were divided into six equal treatment groups and fed 60%, 70%, 80%, 90%, 100% or 110% of their estimated individual SIDLysR. The Lys requirement of each pig was estimated daily using a real-time model. Body composition was measured with dual-energy X-ray densitometry on day 1 and 28 of the experiments. Average daily feed intake increased quadratically (P < 0.05) during both growth phases. Maximum average daily gain (ADG) (0.98 kg) and maximum protein deposition (PD; 170 g/day) were observed in growing pigs fed 100% of the estimated SIDLysR (P < 0.001). During the growing period, PD in BW gain (17% to 19%) and N efficiency (52% to 65%) increased linearly (P < 0.01) with increasing inclusion rates of SID Lys. Finishing pigs had maximum ADG (1.2 kg/day) when they were fed 100% of the requirements. However, the amount of protein in BW gain (13% to 16%) and N efficiency (40% to 55%) increased linearly (P < 0.01) with increasing inclusion rates of SID Lys. In conclusion, the model proposed for precision feeding is correctly calibrated to predict SIDLysR that maximize PD and ADG of average pigs from 25 to 50 kg BW. Still, there is an opportunity to improve the estimation of SIDLysR and N retention in individual pigs by better representing the individual proportion of protein in BW gain and the factors controlling the efficiency of Lys utilization in individual pigs.     

Impact of fibre intake and fibre source on digestibility,gut development,retention time and growth performance of indigenous and exotic pigs

The impact of fibre level and fibre source on digestibility, gastrointestinal tract (GIT) development, total tract mean retention time (MRT) and growth performance was studied in indigenous Mong Cai (MC) and exotic Landrace × Yorkshire (LY) pigs. The diets were based on maize, rice bran, soyabean meal, fish meal and soyabean oil, and cassava residue (CR) or brewer's grain (BG) as fibrous ingredient sources in the high-fibre diets (HF) and were fed ad libitum. A low-fibre diet (LF), containing around 200 g NDF/kg dry matter (DM), was formulated without CR and BG as feed ingredients. The HF diets (HF-CR and HF-BG) were formulated to contain around 270 g NDF/kg DM. The experiment was arranged as a 2 × 3 factorial completely randomized design with six replications, and lasted 27 days. Increased dietary fibre level resulted in a reduction (P < 0.05) in average daily gain, digestibility of organic matter (OM), CP and gross energy (GE) at the ileum and in the total tract, and in MRT, and an increase (P < 0.05) in the feed conversion ratio and in the weight of the GIT (except for small intestine and caecum). The coefficients of total tract digestibility of fibre fractions were higher in HF diets than in the LF diet, with highest values for diet HF-CR, which had a high proportion of soluble non-starch polysaccharides. MC pigs had longer MRT of digesta than LY pigs (P < 0.05), resulting in higher digestibility at the ileum and in the total tract. Across diets and breeds, the total tract apparent digestibility of OM, CP and GE was positively related (R² = 0.80 to 0.84) to the MRT of solids, whereas the MRT was negatively related to the DM intake (R² = 0.60).     

The covalent structure of pig kidney fructose 1,6-bisphosphatase: sequence of the 60-residue NH2-terminal peptide produced by digestion with subtilisin

Digestion of the native pig kidney fructose 1,6-bisphosphatase tetramer with subtilisin cleaves each of the 35,000-molecular-weight subunits to yield two major fragments: the S-subunit (M_{r ca.} 29,000), and the S-peptide (M_r 6,500). The following amino acid sequence has been determined for the S peptide: AcThrAspGlnAlaAlaPheAspThrAsnIle Val ThrLeuThrArgPheValMetGluGlnGlyArgLysAla ArgGlyThrGlyGlu MetThrGlnLeuLeuAsnSerLeuCysThrAlaValLys AlaIleSerThrAla z.sbnd;ValArgLysAlaGlyIleAlaHisLeuTyrGlyIleAla. Comparison of this sequence with that of the NH₂-terminal 60 residues of the enzyme from rabbit liver (El-Dorry et al., 1977, Arch. Biochem. Biophys.182, 763) reveals strong homology with 52 identical positions and absolute identity in sequence from residues 26 to 60.Although subtilisin cleavage of fructose 1,6-bisphosphatase results in diminished sensitivity of the enzyme to AMP inhibition, we have found no AMP inhibition-related amino acid residues in the sequenced S-peptide. The loss of AMP sensitivity that occurs upon pyridoxal-P modification of the enzyme does not result in the modification of lysyl residues in the S-peptide. Neither photoaffinity labeling of fructose 1,6-bisphosphatase with 8-azido-AMP nor modification of the cysteinyl residue proximal to the AMP allosteric site resulted in the modification of residues located in the NH₂-terminal 60-amino acid peptide.     

Biallelic SUN5 Mutations Cause Autosomal-Recessive Acephalic Spermatozoa Syndrome

Acephalic spermatozoa syndrome is a rare and severe form of teratozoospermia characterized by a predominance of headless spermatozoa in the ejaculate. Family clustering and consanguinity suggest a genetic origin; however, causative mutations have yet to be identified. We performed whole-exome sequencing in two unrelated infertile men and subsequent variant filtering identified one homozygous (c.824C>T [p.Thr275Met]) and one compound heterozygous (c.1006C>T [p.Arg356Cys] and c.485T>A [p.Met162Lys]) SUN5 (also named TSARG4) variants. Sanger sequencing of SUN5 in 15 additional unrelated infertile men revealed four compound heterozygous (c.381delA [p.Val128Serfs^∗7] and c.824C>T [p.Thr275Met]; c.381delA [p.Val128Serfs^∗7] and c.781G>A [p.Val261Met]; c.216G>A [p.Trp72^∗] and c.1043A>T [p.Asn348Ile]; c.425+1G>A/c.1043A>T [p.Asn348Ile]) and two homozygous (c.851C>G [p.Ser284^∗]; c.350G>A [p.Gly114Arg]) variants in six individuals. These 10 SUN5 variants were found in 8 of 17 unrelated men, explaining the genetic defect in 47.06% of the affected individuals in our cohort. These variants were absent in 100 fertile population-matched control individuals. SUN5 variants lead to absent, significantly reduced, or truncated SUN5, and certain variants altered SUN5 distribution in the head-tail junction of the sperm. In summary, these results demonstrate that biallelic SUN5 mutations cause male infertility due to autosomal-recessive acephalic spermatozoa syndrome.