Digestibility and metabolism of dietary guanidino acetic acid fed to broilers.

In two feeding experiments the retention of supplemental guanidine acetic acid (GAA) in broilers was investigated. In both experiments, the same three treatments were used; the basal feed was supplemented with 0, 0.6, or 6.0 g GAA per kg of feed. While in a growth study (experiment 1) day-old, male Ross 308 broilers were fed diets for 35 days, these diets were fed for only 8 days to fistulated broilers 34 days of age in a balance study (experiment 2). Feeding 0.6 g/kg GAA did not improve growth performance whereas 6.0 g/kg GAA resulted in a reduction of feed consumption and consequently of weight gain (P ≤ 0.05). Feed conversion was not affected and was 1.48 to 1.49 in all treatments. Increasing levels of dietary GAA gradually increased the creatine concentration in breast muscle and liver tissues (P ≤ 0.05) indicating a transformation and retention of dietary GAA as creatine. In experiment 2 the non-supplemented basal diet allowed us to determine the endogenous GAA, creatine, and creatinine excretions. Accordingly, only small amounts of these metabolites were recovered in feces while they were much higher in urine. Increasing dietary GAA intake increased fecal and renal GAA, creatine, and creatinine excretion and was significant (P ≤ 0.05) at 6.0 g/kg dietary GAA compared to no or 0.6 g/kg GAA supplementation. The mean true fecal digestibility of GAA (99%) was unaffected by the level of supplemental GAA. Considering renal GAA excretions, true availability of supplemental GAA was reduced with increasing dose (83% vs. 71%; P ≤ 0.05). Taking into account creatine and creatinine excretions above those of the basal diet, as they are a consequence of increasing dietary supply, true availability of supplemental GAA shrank from 76% (0.6 g/kg GAA) to 46% (6.0 g/kg GAA; P ≤ 0.05). Changes in blood creatine and creatinine levels reflected the changes observed in the liver and muscle tissues and may suggest increased transport to excretion organs. Data from these experiment were used to estimate the creatine requirement.

Effects of dietary supplementation of meat-type quail breeders with guanidinoacetic acid on their reproductive parameters and progeny performance.

This study was conducted to assess the effect of dietary supplementation of meat-type quail breeders with guanidinoacetic acid (GAA) on their reproductive parameters and progeny performance. Two hundred forty meat-type quails at 25 wk of age were distributed in a completely randomized design with 5 treatments and 8 replicates of 6 birds each. The treatments consisted of 5 dietary levels of GAA (0.00, 0.06, 0.12, 0.18, and 0.24%). The progenies from quail breeders were housed according to breeder treatments and fed a conventional diet based on corn and soybean meal without GAA supplementation. Dietary GAA levels did not affect (P > 0.05) the productivity of meat-type quail breeders, although the concentration of guanidinic compounds (creatine, GAA, and creatinine) in the eggs from the breeders increased linearly (P < 0.05) according to the increase in dietary GAA levels. The number of spermatozoa present in the vitelline membrane was not affected (P > 0.05) by the treatments, but there was a quadratic effect (P < 0.05) of the levels of GAA on fertility, embryonic mortality, and egg hatchability, with the best results estimated at 0.13, 0.15, and 0.14% GAA, respectively. The creatine levels of the pectoral muscle in newborn quails showed a quadratic effect (P ≤ 0.07), and the dietary GAA level of 0.11% was estimated to maximize the muscular creatine level in the progeny. There was a quadratic effect (P < 0.05) of GAA levels on weight gain and feed conversion of progeny at 35 d of age with an optimization point of 0.14% GAA for these variables. Dietary GAA supplementation of meat-type quail breeders increases the availability of creatine in eggs and muscle of progeny, which results in better reproductive parameters and better postnatal progeny performance.

Standardized ileal digestibility of amino acids in alfalfa meal, sugar beet pulp, and wheat bran compared to wheat and protein ingredients for growing pigs.

A total of 11 (8 + 3 for replacement) barrows with an initial BW of 23 kg and fitted with a simple T-cannula at the distal ileum were used in 2 consecutive experiments (Exp. 1 and Exp. 2) to determine the standardized ileal digestibility (SID) of AA in 7 assay feed ingredients according to 2 consecutive duplicated 4 × 4 Latin square designs. In Exp. 1, 3 corn starch-based assay diets were formulated to contain 170 g CP/kg (as-fed basis) from either soybean meal (SBM), canola meal (CM), or meat-and-bone meal (MBM) and 1 assay diet that contained 136 g CP/kg (as-fed basis) from wheat as commonly used feed ingredients for pigs. In Exp. 2, the pigs were fed 4 assay diets formulated to contain 170 g CP/kg (as-fed basis) from either the same SBM as in Exp. 1 or a combination of this SBM and alfalfa meal (AM), sugar beet pulp (SB), or wheat bran (WB) to compare the SID of AA in these feed ingredients with those used in Exp. 1. The SID of AA in CM was lower compared to SBM (P < 0.05) with intermediate values for MBM and wheat. Among fiber rich feed ingredients, SID values were lower in SB compared to WB (P < 0.05) with intermediate values for AM. In AM, SID values ranged between 29 and 45% for Lys, Cys, Thr, and Phe and between 51 and 71% for Arg, His, Ile, Leu, Met, and Val. In SB, SID values ranged between -21 and 46% for Cys, Thr, Phe, and Val and between 51 and 61% for Arg, His, Ile, Leu, Lys, and Met. In WB, SID values were between 55 and 64% for Lys, Cys, Phe, Thr, and Val and between 68 and 80% for Arg, His, Ile, Leu, and Met. The SID values in WB, SB, and AM provided in the present study may improve diet formulation when these feed ingredients are used in diet formulation for pigs.

Standardized ileal digestibilities of crude protein, amino acids, and contents of antinutritional factors, mycotoxins, and isoflavones of European soybean meal imports fed to piglets.

A study with 3 experiments and 3 periods each was conducted to assess the protein value of soybean meal (SBM) batches that were imported into the European Union (EU) from Argentina, Brazil, or the United States (US). Six random SBM batches from each origin were analyzed for contents of CP, AA, ether extract, crude ash, NDF, ADF, oligosaccharides, isoflavones, mycotoxins, trypsin inhibitor activity (TIA), and protein dispersibility index. Piglets were used for determination of standardized ileal digestibilities (SID) of CP and AA in these 18 SBM batches. In each experiment, 12 piglets (initial BW = 5.6 ± 0.7 kg) were surgically fitted with simple T-cannulas at the distal ileum. The piglets were randomly allotted to 18 semisynthetic assay diets, which included 1 of the 18 SBM batches from the 3 origins at an inclusion level of 250 g/kg (as-fed). Average content of CP was 480, 505, and 488 g/kg (as-fed) for Argentinean, Brazilian, and US SBM batches, respectively, and was greater for Brazilian SBM (P ≤ 0.05) compared with the other 2 origins. Contents of most AA were greater (P ≤ 0.05) in Brazilian compared with Argentinean SBM batches. Amino acid contents in US SBM batches ranged between those from Argentina and Brazil. Average TIA were 3.9, 5.1, and 3.4 mg trypsin inhibitor/g CP for Argentinean, Brazilian, and US SBM batches, and were greater (P ≤ 0.05) for Brazilian SBM compared with the other origins. Mycotoxins were detected in 8 out of 18 SBM batches, but all mycotoxin concentrations were less than their critical benchmarks. The contents of individual isoflavones varied considerably and differed (P ≤ 0.05) among SBM origins. The SID of CP, Arg, Phe, Thr, Trp, Asp, Gly, and Ser were greater (P ≤ 0.05) for US compared with Argentinean SBM batches, with intermediate values for Brazilian SBM batches. The obtained SID values were most variable within Argentinean SBM batches and most homogenous within US SBM batches, as indicated by great and small CV, respectively. However, SID of CP and AA were not affected (P > 0.05) by any of the chemical variables measured in this study, according to linear and quadratic regression analyses. Greatest SID values and good homogeneity between individual batches of the same origin were observed for US SBM. The results of this study with piglets will expand the database on SID of CP and AA in SBM from 3 of the major soybean-producing and processing countries.

Supplementation of guanidinoacetic acid to broiler diets: effects on performance, carcass characteristics, meat quality, and energy metabolism.

Creatine, (CREA) a central constituent in energy metabolism, is obtained from dietary animal protein or de novo synthesis from guanidinoacetic acid (GAA). Especially in all-vegetable diets, supplemental CREA or GAA may restore the CREA availability in tissues, and hence, improve performance. In this study, 768 one-d-old male Ross 308 broilers were assigned to 1 of 4 diets: negative control, all-vegetable corn-soybean-based; negative control supplemented with either 0.6 or 1.2 g of GAA per kilogram of feed; and positive control (60, 30, and 30 g/kg of fish meal in the starter, grower, and finisher diets, respectively). Each treatment was replicated in 6 pens of 32 birds each. At the end of the grower period (d 26), 2 birds per pen were euthanized for metabolic measurements. Four broilers per pen were selected at slaughter age (d 39) to determine carcass characteristics and meat quality. Compared with the negative control, GAA supplementation resulted in an improved gain:feed ratio (P < 0.05) and ADG (P < 0.05; + 2.7 and + 2.2% for GAA at 0.6 and 1.2 g/kg, respectively) throughout the entire period. Breast meat yield was higher for the GAA diets compared with that of the negative control birds (P < 0.05; 30.6 vs. 29.4%) and was comparable with that of the positive control birds (30.2%). With regard to meat quality, lower ultimate pH values, higher cooking and press fluid losses, and higher color L* values were observed for the GAA diets compared with those of the negative control diet (P < 0.05). These effects were small, however. The GAA and CREA levels in breast meat were lower and higher, respectively, in GAA-fed birds compared with those of the control birds (P < 0.01). The diets did not affect plasma metabolic traits, except that plasma insulin-like growth factor I concentrations were almost twice as high in animals fed 1.2 g/kg of GAA compared with those of all other treatments. The GAA included in all-vegetable diets improved animal performance for the whole rearing period and increased breast meat yield.

Comparison of standardized ileal amino acid digestibilities in protein supplements and cereal grains for weaned pigs.

Standardized ileal digestibility (SID) of AA in protein ingredients and grains was determined in weaned piglets (5 kg initial BW) using the difference method. Animals were fitted with a simple ileal T-cannula on day 24 or 25 of age. A synthetic diet based on corn (Zea mays) starch and casein was either supplemented with an extruded soybean (Glycine max) meal (ESM), rice (Oryza sativa) protein concentrate (RPC), full fat heat-treated soybeans (SB), corn, barley (Hordeum vulgare), or wheat (Triticum aestivum). The contribution of CP and AA from casein and from the assay feed ingredients to the assay diet averaged 50% each. Each diet was fed to 6 animals according to a row-column design with 3 periods. Apparent ileal digestibility was corrected for basal ileal endogenous losses of AA to obtain SID values. Between RPC and soy products, differences (P < 0.05) in SID of most AA such as Lys, Met, and Thr were greater than within soybean products (ESM and SB). The SID of indispensable AA did not differ (P > 0.05) between barley and wheat, except for Met (P < 0.05). Furthermore, SID of Arg, His, Ile, Phe, and Trp was lower (P < 0.05) in corn than wheat. In conclusion, SID of most AA in the present study with piglets was lower than tabulated SID values determined in studies with grower-finisher pigs. Furthermore, the use of RPC, despite its high CP content, in diets for weaned piglets is limited due to its low SID of AA.

Comparative evaluation of standardized ileal amino acid digestibility in protein supplements for piglets.

Standardized ileal digestibility (SID) of AA was determined in 6 protein ingredients for piglets. A basal diet based on corn (Zea mays) starch and casein was supplemented with fluid-bed-dried porcine intestinal mucosa hydrolysate, spray-dried porcine intestinal mucosa hydrolysate, soy (Glycine max) protein concentrate, 2 batches of soy protein, or full-fat soybeans. The SID of AA did not differ between the 4 soybean products (P > 0.05). Compared to most SID values in the 4 soybean products, SID of AA were lower in the 2 porcine intestinal mucosa hydrolysates (P ≤ 0.05). In conclusion, although the initial trypsin inhibitor contents in the raw soybeans have not been determined, high SID values in the 4 soybean products indicate that the different processing procedures used to manufacture these products were efficient to inactivate trypsin inhibitors. For most AA in the 2 porcine intestinal mucosa hydrolysates, drying procedure did not affect SID of AA, but SID values were generally lower compared to the 4 soybean products.

Effect of soybean meal origin on standardized ileal amino acid digestibility in piglets.

The present study with piglets was conducted to estimate standardized ileal digestibility (SID) of AA in European Union soybean (Glycine max) meal (SBM) imports sourced from Argentina, Brazil, and the United States. Twelve piglets with an average initial BW of 9 kg (47 d of age) were fitted with a simple ileal T-cannula to determine SID of AA in 4 batches of SBM originating either from Argentina (n = 1), Brazil (n = 1), or the United States (n = 2; US1 and US2). A semisynthetic diet based on corn (Zea mays) starch and casein (125 g casein/kg as-fed) was supplemented with 1 of 4 batches SBM at an inclusion rate of 235 g SBM/kg (as-fed) each. The chemical composition did not differ largely, with CP contents ranging from 458 in Argentinean to 461 g/kg (as-fed) in US1 SBM. The SID of indispensable AA ranged from 80% in US2 SBM to 83% in Argentinean SBM and from 74% for Trp in US2 and Brazilian SBM to 91% for Arg in US2, Brazilian, and Argentinean SBM. Most SID values did not differ among the SBM batches (P > 0.05). In conclusion, European SBM imports from Brazil, Argentina, or the United Sates were similar in their chemical composition. For most AA, high and uniform SID values were obtained independent from their source of origin.

Methionine-hydroxy analogue was found to be significantly less bioavailable compared to dl-methionine for protein deposition in growing pigs.

When methionine (Met) is limiting in swine diets, it is commonly supplemented by using anhydrous dl-methionine (DLM, 99% purity) or liquid dl-methionine-hydroxy analogue free acid (MHA-FA, 88% purity). The objective of this experiment was to test the null hypothesis that the bioavailability of DLM and MHA-FA were not different for growing pigs, using the indicator amino acid (AA) (phenylalanine, Phe) oxidation (IAAO) method in a slope-ratio assay. Six barrows (mean BW during study: 21.1 kg) received seven dietary treatments with all pigs receiving all diets in random order at an intake of 95 g/kg BW0.75. The basal diet (BD) contained analyzed content of 15.1% CP, 0.20% Met, 0.73% Phe and all other AA in excess of requirement. The BD was supplemented with three graded levels of DLM or MHA-FA on an equimolar basis. Dietary treatments only varied in Met content and included: (i) BD, (ii) BD + 0.034% DLM, (iii) BD + 0.054% DLM, (iv) BD + 0.086% DLM, (v) BD + 0.029% MHA-FA, (vi) BD + 0.078% MHA-FA and (vii) BD + 0.107% MHA-FA, as analyzed. Indicator AA oxidation was determined during 4 h studies, where pigs were fed half-hourly meals each equal to 1/32 of their daily feed allowance. Each meal was mixed with 258.7 kBq (s.e. 2.6) of l-[1-14C]Phe with a prime of 3.5 times the half-hourly dose added to the first meal. The slope of the decrease in IAAO calculated by linear regression analysis was greater (P = 0.012) for DLM supplementation (9.87 ± 1.450 per g, 1.488 ± 0.215% per mmol) than for MHA-FA (6.48 ± 0.89 per g, 1.107 ± 0.152% per mmol). The ratio of slopes indicated a bioavailability of MHA-FA on a product basis, relative to DLM, of 65.7%. Bioavailability on an equimolar Met basis, calculated from the ratio of the slopes was 74.4% for MHA-FA, relative to DLM. In conclusion, these results indicate that the metabolic bioavailability of MHA-FA for growing pigs is appreciably lower than that of DLM on both an equimolar and a product basis.

Effect of dietary protein level on growth performance, indicators of enteric health, and gastrointestinal microbial ecology of weaned pigs induced with postweaning colibacillosis.

The effect of dietary CP level on performance, enteric health, and gastrointestinal microbial ecology of weaned pigs challenged with enterotoxigenic Escherichia coli (ETEC) K88 was investigated in a 14-d study. Forty weaned pigs (BW = 5.32 +/- 0.24 kg, mean +/- SD), housed 4 per pen, were randomly assigned to 2 diets (5 pens/diet): 1) 22.5% CP or 2) 17.6% CP supplemented with AA. Diets contained the same amount of ME and standardized ileal digestible Lys, Met + Cys, Thr, and Trp based on the ideal protein ratio. Isoleucine and Val were added to the 17.6% CP diet up to the level in the 22.5% CP diet. On d 8 postweaning, pigs were challenged with 6 mL of ETEC suspension (10(10) cfu/mL) by gavage. Feed disappearance and BW were measured on d 7, 9, 10, 12, and 14 for determination of ADG, ADFI, and G:F. One pig from each pen was serially slaughtered on -1, 3, and 7 d postchallenge (10 pigs/d of slaughter) to evaluate gut morphology, and gut microbial ecology and metabolites. Pigs fed the 22.5% CP diet had greater (P < 0.05) ADG and G:F than those fed the 17.6% CP diet before infection, but performance was similar between the 2 diets after ETEC challenge and overall. On d 3 after challenge, ETEC was not detected in the ileal digesta of pigs fed the 17.6% CP diet but was detected in the ileal digesta of 80% of pigs fed the 22.5% CP diet (5.22 +/- 1.07 cfu/g, mean +/- SD). Pigs fed the 17.6% CP diet had a greater (P < 0.01) prevalence of order Clostridiales (73 vs. 50%), family Lachnospiraceae (43 vs. 18%), and genus Roseburia (13 vs. 3%) in the colon digesta 7 d after challenge compared with those fed the 22.5% CP diet. The richness and diversity of bacteria in the colon digesta were less (P < 0.05) in pigs fed the 17.6% CP diet than in those fed the 22.5% CP diet at -1, 3, and 7 d postchallenge. Pigs fed the 22.5% CP diet had greater (P < 0.05) ammonia N concentration in the colon digesta on -1 and 7 d after challenge compared with those fed the 17.6% CP diet. Pigs fed the 22.5% CP diet had deeper (P < 0.05) crypts 1 d before challenge, shorter villi 3 d after challenge, and reduced villus height:crypt depth 1 d before and 3 d after challenge compared with those fed the 17.6% CP diet. In conclusion, a reduction in the dietary CP level of weaned pigs from 22.5 to 17.6% with AA supplementation impaired growth performance before, but not after, the ETEC challenge and increased the relative composition of butyrate producing bacteria in the colon digesta after ETEC challenge.

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.

Divergent modulation of swine ileal microbiota by formic acid and methionine hydroxy analogue-free acid.

Management of intestinal microbiota of monogastric animals has increased in importance since the ban of growth promoting antibiotics in many countries. Organic acids have been used as alternatives to antibiotics by many feed manufacturers. Regardless of the wide usage, the effect, dose response and mode of action of acids on intestinal microbes is poorly understood. In this study, we investigated the effects of dietary supplementation of three commonly used products, namely formic acid (FA) (90%), dl-methionine (DLM) (99%) and liquid methionine hydroxy analogue-free acid (88%), on ileal microbiota of pigs. Laboratory simulation system, mimicking swine ileum, was used to study the products at various concentrations and combinations. Furthermore, selected combinations were tested in a piglet trial to confirm the findings made in in vitro studies. FA turned out to have a dual effect on ileal microbiota. At concentrations below 0.5%, it significantly stimulated bacteria, but at higher inclusion rates it was highly inhibitory. This finding, which was consistent in in vitro and in vivo studies, implies that reducing the dose of FA does not lead to a diluted inhibitory effect, but in fact, an opposite, stimulatory effect on intestinal microbiota. It is highly important that feed compounders acknowledge this finding. Unlike FA, the inhibitory effect of methionine hydroxy analogue on ileal bacteria was linearly dose dependent and significant at inclusion levels above 0.2%, in vitro. Partial replacement of methionine hydroxy analogue by FA, or FA by methionine hydroxy analogue, led to an unpredictable outcome due to the dual effects of FA; e.g., a minor inclusion of added FA changed the inhibitory effect of methionine hydroxy analogue into microbial stimulation by FA. Inhibition of ileal microbiota by methionine hydroxy analogue was detected only in in vitro studies, suggesting that intact methionine hydroxy analogue may not have reached the ileum, in live animals. Therefore, if the target is to ensure the inhibitory effect of FA, the FA level in feed should be kept above 0.6%, and not reduced, if methionine hydroxy analogue is used as a methionine source instead of DLM. DLM was totally inert with regard to bacterial growth and metabolism, both in vitro and in vivo. The results of these studies reveal the importance of knowing how each acid product works. Inconsistent results in animal trials may have been partly due to quadratic dose-response effects of products, and unpredictable product combination effects.

Apparent absorption of methionine and 2-hydroxy-4-methylthiobutanoic acid from gastrointestinal tract of conventional and gnotobiotic pigs.

The effect of commensal microbiota and feeding corn or wheat/barley-based diets on the apparent gastrointestinal absorption of dl-methionine (MET) and 2-hydroxy-4-methylthiobutanoic acid (MHA-FA) was studied in conventional (n = 32) and gnotobiotic pigs (n = 24). Conventional pigs (CON) were vaginally delivered and sow-reared until weaning at 14 days of age. Gnotobiotic pigs were derived by caesarian section and reared in HEPA (high efficiency particulate air)-filtered isolator units with ad libitum access to a milk-based formula. Corn or wheat/barley-based diets were fed to all pigs from 14 to 24 days of age. At 24 days of age, after an overnight fast, pigs were fed 20 g/kg BW of experimental diet supplemented with 107 Bq of either 3H-l-MET or 3H-l-MHA-FA per kg of feed and chromic oxide (0.5% wt/wt). Pigs were killed for sample collection 3 h after consuming the meal. Residual 3H-MET and 3H-MHA-FA were estimated in gastrointestinal contents as the ratio of 3H : chromic oxide in digesta samples to the ratio of 3H : chromic oxide in feed. In CON pigs, feeding a wheat/barley-based diet increased (P < 0.05) total aerobes, whereas supplementation with MHA-FA increased (P < 0.05) total aerobes and lactobacilli populations in proximal small intestine (SI). Among the gnotobiotic pigs, bacterial contamination occurred such that eight pigs (two isolators) were monoassociated with a Gram-negative bacteria closely related to Providencia spp. and 16 pigs (four isolators) were monoassociated with Gram positive Enterococcus faecium. Species of monoassociated bacterial contaminant and diet composition did not affect residual methionine or MHA-FA in digesta. In both CON and monoassociated (MA) pigs, methionine and MHA-FA were retained in stomach (92%) but disappeared rapidly from proximal SI. Residual methionine and MHA-FA in digesta was not different in MA pigs; however, in CON pigs, less (P < 0.01) apparent residual methionine was found in digesta recovered at 25% (from cranial to caudal) and 75% of SI length compared with MHA-FA. Apparent residual methionine was 16% and 8% compared with 34% and 15% for MHA-FA, at the 25% and 75% locations, respectively. In proximal SI tissue, significantly (P < 0.05) higher radioactivity (cpm/mg wet tissue) was associated with MET pigs (8.56 ± 0.47) as compared to MHA-FA ones (5.45 ± 0.50). This study suggests that microbial metabolism of MHA-FA increases retention in small intestinal digesta relative to methionine and contributes, in part, to the lower bioefficacy of MHA-FA compared to methionine.

Effect of dietary crude protein level on basal ileal endogenous losses and standardized ileal digestibilities of crude protein and amino acids in newly weaned pigs.

The study was carried out to estimate basal ileal endogenous crude protein and amino acid losses (IAAL(B)) and standardized ileal digestibilities (SID) of crude protein (CP) and amino acids (AA) originating from casein in newly weaned pigs from linear relationships between ileal recoveries and dietary intakes of graded levels of CP and AA. A total of 14 (12 + 2 for replacement) 3-week-old barrows weighing 5.7 kg was fitted with simple T cannulas at the distal ileum. At 28 days of age, the pigs were randomly allocated to the six experimental diets with two pigs per diet in four weekly repeated measurements. Corn starch-based diets, containing six graded levels of CP from casein (90, 155, 220, 285, 350, 415 g/kg CP as-fed basis respectively), were formulated. Titanium dioxide (TiO(2)) was included as a digestibility marker. Each experimental period consisted of 7 days. Ileal digesta were collected for a total of 24 h during day 6 and 8. Splitting the total range of 90 to 415 g/kg CP into smaller ranges, i.e. from 90 to 220, 220 to 350 or 285 to 415 g/kg CP, provides estimates for CP and AA recoveries which are not significantly different from zero. As a result, dietary CP and AA originating from casein are completely digested and absorbed until the end of the small intestine. In addition, the use of large ranges of dietary CP levels showed that IAAL(B) were affected (p < 0.050) by the dietary CP content. Accepting that ileal recoveries of CP and AA are exclusively of endogenous origin when purified corn starch casein-based diets are fed, IAAL(B) were estimated as a function of the dietary CP level. There were linear increases (p < 0.050) in IAAL(B) when the dietary CP content was increased from 90 to 415 g/kg. Average IAAL(B) expressed in g/kg dry matter intake (DMI) were 16.3, 0.7, 0.2, 0.9 and 0.2 for CP, lysine, methionine, threonine and tryptophan respectively. These values are in close agreement with IAAL(B) obtained in grower-finisher pigs.

An estimate of the methionine requirement and its variability in growing pigs using the indicator amino acid oxidation technique.

Although AA requirements for the mean of a population of growing pigs have been established using traditional methods, there are no estimates of the variability within the population and whether this variation differs among AA. With the increased use of supplemental Lys in pig diets, there will be an increased need to supplement Met, commonly the second or third limiting AA in corn-soybean diets. The indicator AA oxidation method allows repeated measurements in a short period of time so that the AA requirement can be determined for individual pigs at a similar physiological stage. The objective of this study was to determine the mean Met requirement in individual gilts and to estimate the related variability. Six individually housed female pigs (initial BW = 8.8 kg, SD 1.5) each received diets providing 6 levels of dl-Met. The isonitrogenous and isoenergetic diets contained 0.187, 0.250, 0.290, 0.320, 0.350, and 0.377% Met (analyzed, as-fed basis). Cysteine (0.48%) and Lys (1.44%) concentrations were similar for all diets. Pigs were adapted for 6 d to the basal corn-soybean meal diet (0.187% Met), which was offered at 95 g/kg(0.75) of BW to ensure complete consumption of the test diets. During 4-h oxidation studies, 313.4 kBq, (SD 35.6) of L-[1-(14)C]Phe was mixed with each of 8 half-hourly meals, and expired CO(2) was collected. The breakpoint in Phe oxidation, representing the Met requirement, and its variability, was determined using 2-phase linear regression. Phenylalanine oxidation decreased as the Met content increased from 0.187 to 0.29%. Phenylalanine oxidation was not different (P > 0.2) for diets ranging from 0.320 to 0.377% Met. The dietary Met requirement varied from 0.320 to 0.373% for individual pigs. The mean Met requirement for individual pigs was determined to be 0.340% of diet (SD = 0.024%, CV= 7.1%), with 0.340, 0.364, and 0.388% covering the requirement of 50, 66, and 95% of the population, respectively. The present mean population estimate was similar to the recommended dietary Met concentration of 0.325% for pigs of this BW and feed intake. To maximize profitability, Met levels in starter pig diets should be determined, depending on the cost of crystalline Met and the fraction of the population whose requirement is to be met.

Effect of low-protein amino acid-supplemented diets on the growth performance, gut morphology, organ weights and digesta characteristics of weaned pigs.

A 21-day study was conducted to determine whether isoleucine might limit the performance of piglets fed low-crude protein (CP), amino acid (AA)-supplemented diets and to investigate the potential benefits of low-CP diets on gastrointestinal health in weaned pigs. Ninety-six piglets (initial BW = 6.44 ± 0.14 kg), housed four per pen, were randomly assigned to one of four diets, resulting in six replicate pens per diet. Dietary treatments were as follows: (1) 210 g/kg CP diet, (2) 190 g/kg CP diet deficient in isoleucine, (3) 190 g/kg CP diet supplemented with crystalline isoleucine up to the level in the 210 g/kg CP diet and (4) 170 g/kg CP diet supplemented with isoleucine and valine on the ideal protein ratio basis (60% and 70% relative to lysine, respectively). Pigs were allowed to adapt to the new environment for 4 days before the experiment commenced. Overall, pigs fed the 210 g/kg CP diet had higher (P < 0.05) average daily gain and lower (P < 0.05) feed : gain ratio compared with those fed the other diets. The faecal consistency score of pigs fed the 210 g/kg CP diet was higher (P < 0.05) than those fed the other diets. Pigs fed the 170 g/kg diet had lower (P=0.02) small intestine weight than those fed the 210 g/kg CP diet. Pigs fed the 210 g/kg CP diet had deeper (P < 0.05) crypt in the duodenum and ileum and higher (P < 0.05) ammonia N concentration in caecal digesta than those fed the other diets. There were no effects of diet on microbial population and volatile fatty acid concentration in the caecal digesta except for propionic acid whose concentration was higher (P < 0.05) for pigs fed the 170 g/kg diet than those fed the 190+isoleucine and the 210 g/kg CP diets. The results indicate that the low-CP, AA-supplemented diet reduced crypt hypertrophy, ammonia N concentration in the caecal digesta, small intestine weight and the performance of piglets. Also, the results of the current study were inconclusive with respect to whether isoleucine may limit the performance of pigs fed a low-CP, AA-supplemented diet.

Effect of dietary protein content on ileal amino acid digestibility, growth performance, and formation of microbial metabolites in ileal and cecal digesta of early-weaned pigs.

Diarrhea incidence in weaned pigs may be associated with the concentration of intestinal microbial metabolites (ammonia, amines, and VFA) that are influenced by dietary CP content. Three experiments were conducted to determine effects of a low-protein, AA-supplemented diet on ileal AA digestibility, growth performance, diarrhea incidence, and concentration of microbial metabolites in ileal and cecal digesta of pigs weaned at 14 d of age. In Exp. 1, 8 pigs fitted with a simple T-cannula at the distal ileum were assigned in a crossover design to 2 diets containing 24 or 20% CP using wheat, corn, full-fat soybeans, whey powder, fish meal, and blood plasma as the main ingredients. Supplemental AA were added to the diets to meet the AA standards according to the 1998 NRC recommendations. Chromic oxide was used as an indigestible marker. Diets were fed at 2.5 times the ME requirement for maintenance. The reduction of dietary CP decreased (P < 0.05) the apparent ileal digestibility of most AA, except Lys, Met, Thr, Val, and Pro. Dietary CP content did not affect the pH of ileal digesta or ileal concentrations of ammonia N, cadaverine, putrescine, or VFA. In Exp. 2, 8 pigs fitted with a simple T-cannula in the cecum were assigned to 2 diets, similar to Exp. 1. Dietary CP content did not affect the pH of cecal digesta. The reduction in CP content decreased (P < 0.05) cecal ammonia N, acetic acid, isobutyric acid, isovaleric acid, total VFA, and putrescine concentrations by 28 to 39%. In Exp. 3, 32 pigs were assigned to 2 diets, similar to Exp. 1, according to a randomized complete block design. Pigs had free access to feed and water. Dietary CP content did not affect growth performance or fecal consistency scores during the 3-wk study, and diarrhea was not observed. The results of these experiments indicate that lowering the dietary CP content combined with supplementation of AA markedly reduced the production of potentially harmful microbial metabolites in cecal digesta of early-weaned pigs without affecting growth performance.

Efficacy of DL-methionine hydroxy analog free acid and DL-methionine as methionine sources for pigs.

Two experiments were conducted to evaluate the efficacy of dietary DL-methionine hydroxy analog-free acid (MHA-FA, 88%) compared with DL-methionine (DLM, 99%) as Met sources in pigs. In Exp. 1, a total of 245 crossbred pigs (initial BW of 6.4 kg [SD = 0.5]) were allotted to 7 treatments in 7 replicates for an experimental period of 28 d. The basal diet (BD) was formulated to contain 17.5% CP and 0.21% Met. Dietary treatments included 1) BD, 2) BD + 0.030% DLM, 3) BD + 0.060% DLM, 4) BD + 0.090% DLM, 5) BD + 0.034% MHA-FA, 6) BD + 0.068% MHA-FA, and 7) BD + 0.103% MHA-FA; the MHA-FA was supplemented on an equimolar basis to the DLM. Because of a nonlinear response, exponential regression analysis was used to evaluate the responses, and a comparison of the equations was then made to determine the relative effectiveness of the 2 Met sources. With increases in dietary Met, weight gain increased (P < 0.05). Compared with DLM on a product-to-product (wt/wt) basis, the relative effectiveness of MHA-FA was calculated to be 73% for increasing weight gain and 54% for decreasing the feed:gain. In Exp. 2, a total of 30 weanling barrows [initial BW of 16.8 kg (SD = 2.8)] were used in a metabolism study to evaluate the relative value of MHA-FA to DLM. The BD was formulated to contain 16.9% CP and 0.21% Met. Dietary treatments included 1) BD, 2) BD + 0.030% DLM, 3) BD + 0.060% DLM, 4) BD + 0.046% MHA-FA, and 5) BD + 0.092% MHA-FA; the MHA-FA levels were chosen based on a pre-experiment estimate of bioequivalence in an attempt to provide approximately equal pig responses. There was no difference in fecal N output among the treatments; however, urine N linearly decreased with increasing concentrations of both sources (P = 0.034 for DLM, and P = 0.007 for MHA-FA), which resulted in a linear increase in retained N for both DLM (P = 0.012) and MHA-FA (P = 0.005). In addition, N retention (% of intake) linearly increased with increasing level of DLM (P = 0.014) and MHA-FA (P = 0.007). Using a slope-ratio procedure for comparison of the responses from the 2 sources, the relative biological equivalence value of MHA-FA to DLM in this experiment was 64.2% based on percent N retention and 66.3% based on the grams of N retained per day. Based on the results from both experiments, these data indicated that the mean relative bioequivalence of MHA-FA to DLM was 64% on a product-to-product (wt/wt) basis or 73% on an equimolar basis.

Performance responses and indicators of gastrointestinal health in early-weaned pigs fed low-protein amino acid-supplemented diets.

The effects of low-protein AA-supplemented diets on piglet performance, visceral organ mass, incidence of diarrhea, intestinal microbial population, and fermentation were studied in a 3-wk trial. After a 7-d adaptation period, 96 piglets (approximately 6.2 kg of initial BW) were assigned to 4 corn-wheat, soybean meal-based dietary treatments in a completely randomized design to give 6 replicate pens per treatment (n = 4 piglets per pen). The treatments were a control wheat-corn-soybean meal-based phase I diet containing 23% CP, or the same diet with CP reduced to 21%, 19%, or 17% and supplemented with crystalline AA to achieve equal standardized ileal digestible contents of Lys, Met plus Cys, Thr, and Trp in all diets. Diets were formulated to similar nutrient levels and provided ad libitum. Blood from all pigs was taken on d 0, 7, 14, and 21 for determining plasma urea N. Weekly feed intake, BW changes, and G:F were determined. On d 21, 2 pigs per pen were randomly selected and killed to determine small intestinal morphology, digesta pH and ammonia levels, and luminal microbial counts. Average daily feed intake, ADG, and G:F were not affected (P > 0.10) by reducing CP to 21%, but a reduction to 19% or 17% decreased ADFI (P < 0.001) and ADG (linear, P < 0.001; quadratic, P < 0.05) over the 3-wk study period. Reducing CP to 19% had no effect (P > 0.10) on G:F; however, this response criterion was decreased linearly (P < 0.001) over the 3-wk study period as dietary CP declined. Water usage was only numerically decreased (P > 0.10) with dietary CP reduction. Plasma urea N was decreased linearly (P < 0.01) with CP reduction. Reducing CP from 23 to 17% had a linear (P < 0.05) and cubic effect on stomach and liver weights, respectively. Although histological data showed some differences among diets, no distinct trend was evident. Ammonia N in ileal digesta was reduced linearly (P < 0.01) as dietary CP was decreased. With the exception of valeric acid, VFA levels in ileal digesta of piglets fed low-protein diets were generally lower (P < 0.05) compared with the control diet. Diet had no effect on intestinal microbial counts (P > 0.10). The results show that piglet performance may suffer when dietary CP is reduced by 4 or more percentage units from 23% and support the hypothesis that low-CP diets help maintain enteric health in pigs by lowering toxic microbial metabolites such as ammonia.

Efficacy and cognitive side effects of tiagabine and topiramate in patients with epilepsy.

OBJECTIVE: Whereas the efficacy of the newer antiepileptic drugs (AEDs) is well established, there remain questions regarding their cognitive side effects. Therefore, we performed a comparative open randomized trial with TPM and TGB as add-on therapy, with particular consideration of cognition, mood, and health-related quality of life (HRQOL). METHODS: Forty-one patients with refractory epilepsy were randomly assigned to one of the two treatment groups (TPM vs TGB) and received neuropsychological testing at baseline (T1), after titration (3 months, T2), and during the maintenance phase (another 3 months, T3). Tests included measures of intelligence, attention, working memory, episodic memory, language, and self-report questionnaires regarding mood and HRQOL. Twenty patients (8 TPM, 12 TGB) discontinued the trial for different reasons (no group difference). RESULTS: Seizure outcome (intention-to-treat analysis) was comparably good in both groups (8.1% seizure free, 29.7% seizure reduction>50%). From baseline to after the titration paired sample t tests revealed significant deterioration in verbal fluency, language comprehension, working memory, and visual block tapping under TPM and a deterioration in verbal memory (delayed free recall) in the TGB group. These functions remained stable in the maintenance phase. Self-report measures initially indicated concerns about AED side effects in both groups and concerns about worse cognitive functioning and depression under TPM. In the maintenance phase the TGB group reported feeling a lack of energy, whereas patients on TPM demonstrated improvement on all QOLIE scales on a descriptive level. CONCLUSION: This study demonstrates the comparable efficacy of TPM and TGB. Consistent with previous reports, TPM but not TGB appears to be associated with persistent negative cognitive side effects on frontal lobe-associated functions, the degree of which may be estimated by the fact that this effect was observed with a very small sample size. In contrast, in patients taking TPM, initially negatively affected HRQOL returns to baseline in the long run on a descriptive level. The latter finding may be interpreted in accordance with the observation that objective performance and subjective self-report under TPM can be dissociated.