Effect of carbohydrase and protease on growth performance and gut health of young broilers fed diets containing rye, wheat, and feather meal.

An experiment was conducted to characterize a gut health challenge model consisting of a diet containing rye, wheat, and feather meal and a mild mixed-species Eimeria challenge, and to evaluate the effect of carbohydrase and protease on growth performance and gut health of young broilers. The study included 4 treatments: negative control, carbohydrase alone, protease alone, and combination of carbohydrase and protease. Each test diet was fed to 18 battery pens of broilers with 8 male birds per pen from 0 to 22 d of age. Carbohydrase improved body weight, feed intake, and feed conversion ratio (FCR) on d 7, 14, and 21(P < 0.01). Protease increased body weight on d 7 and 21 and improved 0 to 7 d FCR (P < 0.05). More lymphocyte infiltration was observed in small intestine mucosa of negative control birds on d 8, carbohydrase supplementation lessened this. Both carbohydrase and protease reduced digesta viscosity on d 22 with the carbohydrase effect being the greater of the two, and the combination effect was not different from the carbohydrase effect alone (P < 0.01). Ileal Clostridium perfringens of 15-day-old broilers was decreased by carbohydrase, a further reduction was achieved by combining carbohydrase with protease (P = 0.01). Liver vitamin E concentration on d 15 (P < 0.01) and 22 (P = 0.02) was increased by carbohydrase, and the carbohydrase effect was greater in the presence of protease on d 22 (P = 0.04). Plasma α-1-acid glycoprotein level and liver Zn and Cu concentrations of broilers were reduced by carbohydrase on d 15 (P < 0.01). Broilers fed carbohydrase had higher levels of plasma zeaxanthin on d 22 and higher levels of plasma lutein on d 15 and 22 (P < 0.01). In summary, a rye wheat based diet containing feather meal when fed to broilers in addition to a mild Eimeria challenge induced subclinical enteritis characterized by digestion inefficiency, dysbacteriosis, inflammation, and gut barrier failure; carbohydrase and protease could be effective tools to improve growth performance and gut health of broilers suffering from this type of subclinical enteritis.

Supplementing antioxidants to pigs fed diets high in oxidants: II. Effects on carcass characteristics, meat quality, and fatty acid profile.

The study was conducted to determine effects of dietary supplementation with a blend of antioxidants (ethoxyquin and propyl gallate) on carcass characteristics, meat quality, and fatty acid profile in finishing pigs fed a diet high in oxidants. A total of 100 crossbred barrows (10.9±1.4 kg BW, 36±2 d of age) were randomly allotted to 5 diet treatments (5 replicate pens per treatment, 4 pigs per pen). Treatments included: 1) HO: high oxidant diet containing 5% oxidized soy oil and 10% PUFA source which contributed 5.56% crude fat and 2.05% docosahexanoic acid (DHA) to the diet; 2) VE: the HO diet with 11 IU/kg of added vitamin E; 3) AOX: the HO diet with antioxidant blend (135 mg/kg); 4) VE+AOX: the HO diet with both vitamin E and antioxidant blend; and 5) SC: a standard corn-soy control diet with nonoxidized oil and no PUFA source. The trial lasted for 118 d; on d 83, the HO diet pigs were switched to the SC diet due to very poor health. From that point, the VE pigs displayed the poorest performance. On d 118, 2 pigs from each pen were harvested for sampling. Compared to pigs fed SC diet, the HO and VE pigs (P<0.05) showed lighter carcass weight, less back fat, less lean body mass, and smaller loin eye area. In addition, the VE pigs had decreased dressing percentage than the AOX and VE+AOX pigs (65.7 vs. 75.3 and 74.2%). Compared to the SC pigs, greater moisture percentage (74.7 vs. 77.4%) and less extractable lipid content (2.43 vs. 0.95%) were found in VE fed pigs (P<0.05). Drip loss of loin muscle in VE pigs was less than SC pigs (0.46 vs. 3.98%, P=0.02), which was associated with a trend for a greater 24-h muscle pH (5.74 vs. 5.54, P=0.07). The antioxidant blend addition in the high oxidant diet attenuated all of these effects to levels similar to SC (P>0.05), except a* value (redness) and belly firmness. Visible yellow coloration of backfat and lipofuscin in HO and VE pigs was observed at harvest at d 118. The high oxidant diet resulted in greater concentration of DHA in backfat (P<0.001); switching the diet on d 83 resulted in HO pigs having a similar fatty acid profile to SC at d 118 pigs. Vitamin E concentration in plasma and muscle was greater in HO and SC than VE, AOX, and VE+AOX on d 118. Feeding the high oxidant diet caused a series of changes in carcass characteristics and meat quality. Addition of antioxidant blend attenuated many of these, whereas the protective effects of supplemental vitamin E at 11 IU/kg were minimal during the finisher phase of the study.

Organic trace minerals and 25-hydroxycholecalciferol affect performance characteristics, leg abnormalities, and biomechanical properties of leg bones of turkeys.

Leg problems and resulting mortality can exceed 1% per week in turkey toms starting at approximately 15 wk of age. Dietary supplementation of organic trace minerals (MIN) and 25-hydroxycholecalciferol (HyD) may improve performance, decrease incidence of leg abnormalities, and increase bone strength. Nicholas 85X700 toms were assigned to 4 treatments consisting of a factorial arrangement of 2 concentrations of MIN (0 and 0.1% of Mintrex P(Se), which adds 40, 40, 20, and 0.3 mg/kg of Zn, Mn, Cu, and Se, respectively) and 2 concentrations of HyD (0 and 92 microg/kg of HyD). Diets were formulated to be equal in nutrient content and fed ad libitum as 8 feed phases. Feed intake and BW were measured at 6, 12, 15, 17, and 20 wk of age. Valgus, varus, and shaky leg defects were determined at 12, 15, 17, and 20 wk of age. Tibia and femur biomechanical properties were evaluated by torsion and bending tests at 17 wk of age. There were no treatment effects on BW. Only MIN significantly improved feed conversion ratio through to 20 wk of age. Cumulative mortality at 3 wk of age was greater among the MIN birds, but it was lower by 20 wk (P = 0.085). The MIN decreased the incidence of varus defects at 17 wk of age; shaky leg at 12, 15, and 17 wk of age; and valgus defects at 15, 17, and 20 wk of age. There were no MIN x HyD interaction effects on individual gait problems. Maximum load and the bending stress required for tibias to break in a 4-point assay were increased with MIN supplementation, especially when HyD was also added. Maximum shear stress at failure of femoral bones in a torsion assay was increased by supplementation with both MIN and HyD together. Dietary supplementation of MIN and HyD may improve biomechanical properties of bones. Dietary MIN supplementation may improve feed conversion of turkeys, likely by decreasing leg problems.

Determining the methionine activity of Mintrex organic trace minerals in broiler chicks by using radiolabel tracing or growth assay.

Mintrex Zn, Mintrex Cu, and Mintrex Mn organic trace minerals contain 16% Zn, 15% Cu, and 13% Mn with 80, 78, and 76% 2-hydroxy-4-(methylthio)butanoic acid (HMTBA) by weight as the organic ligand, respectively. Our objective was to determine if HMTBA from Mintrex was fully available as a Met source. In experiment 1, thirty-six broilers (7 to 10 d old) were orally gavaged with methyl-(14)C-labeled HMTBA, either as free HMTBA (Alimet feed supplement) or Zn bis(-2-hydroxy-4-methylthiobutyrate) (Mintrex Zn). Radiolabel incorporation from either source into protein was measured as a marker of bioavailable Met activity. Results demonstrated that the HMTBA from Mintrex Zn was equally available as free HMTBA to support protein synthesis. In experiment 2, five hundred seventy-six 1-d-old broilers were allotted to 12 dietary treatments (TRT) for a 21-d growth assay. A TSAA-deficient diet containing 0.70% total TSAA (TRT 1) was supplemented with 0.05, 0.10, 0.15, and 0.20% free HMTBA (TRT 2 to 5) to establish the standard Met response curve. Treatment 6 was analogous to TRT 2 but had an additional 160 ppm Zn, 80 ppm Cu, and 160 ppm Mn as sulfates. Treatments 7 to 12 were identical to TRT 2 but supplemented with 40 or 160 ppm Zn from Mintrex Zn, 20 or 80 ppm Cu from Mintrex Cu, or 40 or 160 ppm Mn from Mintrex Mn, respectively. For TRT 1 through 6, growth performance increased due to increasing Met addition (P < 0.01) but not to increasing inorganic trace minerals. For Mintrex Zn, Cu, and Mn (TRT 7 to 12), there was a linear increase in cumulative gain:feed ratio (P < 0.04), and for Mintrex Zn and Mn, there was a linear increase in cumulative gain (P < 0.03) to increasing Mintrex addition. A 1-slope broken-line model was used to calculate bioavailable Met activity from Mintrex for comparison with actual intake values. Results indicated that HMTBA from Mintrex was fully available as a Met source.

Absorption and digestive tract metabolism of 2-hydroxy-4-methylthiobutanoic acid in lambs.

Anabolic availability of the hydroxyl methionine analog, 2-hydroxy-4-methylthiobutanoic acid (HMTBA), given as oral doses to lambs, was quantified both directly as appearance in the portal vein and as synthesis to Met by digestive tract tissues. Eight lambs, prepared with vascular catheters in the mesenteric and portal veins plus the aorta, received twice daily for 7 d either 0.46 g or 2 g of HMTBA. On d 7, [1-13C]HMTBA was supplied as 1 oral dose while [methyl-2H3]Met was infused into the jugular vein. Peak absorption as HMTBA occurred 70 to 90 min after the oral dose. All digestive tract tissues converted HMTBA to Met, equivalent to 24% of the Met provided by the diet for the larger HMTBA dose. Overall, total availability of HMBTA averaged 17.9% of the dose (range 10.6 to 27.9%), with 12.5% (range 7 to 22%) as absorbed HMBTA and the remainder as Met synthesized by digestive tract tissues. Release of 13CO2 into the portal vein accounted for another 23% of the dose. In all digestive tract tissues, the d-isomer was present in a smaller proportion than in the dose. In terms of whole-body kinetics, HMTBA loss from the plasma followed first-order kinetics, with a mean biological half-life of 76 min. Using this value, a simple model was devised to estimate HMTBA absorption based on peripheral plasma samples. When compared with direct measures of absorption, the model gave a slope of 0.81 (R2 = 0.68) and offers a practical means to test HMTBA availability to animals.

Absorption of 2-hydroxy-4-methylthiobutyrate and conversion to methionine in lambs.

Absorption and metabolism of the Met hydroxy analog 2-hydroxy-4-methylthiobutyrate (HMTBA) was examined using stable isotopes. In the first trial, Dl[1-13C]HMTBA was infused for 6 h (7.4 micromol/min) into the abomasum, and [2H3]Met was infused into the mesenteric vein, of 4 lambs prepared with vascular catheters across the splanchnic bed. Daily, lambs were offered 35 g of a mixed forage-concentrate feed/kg. Recovery of HMTBA at the portal vein was 87%, and of this, 63% bypassed the liver. In contrast, hepatic extraction of Met equaled or exceeded net absorption. Only small quantities of Met synthesized from HMTBA were exported from either the digestive tract or liver, but there was substantial and significant input from posthepatic tissues. In a second experiment, 3 of the lambs were killed following 4-h infusions of DL[1-13C]HMTBA and [2H3]Met with enrichments monitored in 15 tissues. Only kidney showed [1-13C]Met enrichment higher than plasma, which suggests that it must be a primary source of plasma Met derived from HMTBA. Based on comparison of plasma and intracellular [1-13C]:[2H3]Met enrichments, all tissues synthesized Met from HMTBA but to significantly different extents. The lowest values were for muscle, skin, brain, and lung; intermediate conversions occurred in rumen, omasum, abomasum, duodenum, jejunum, ileum, and cecum; and the greatest synthesis, equivalent to 22 to 24% of Met entry into cells, was observed for liver and kidney. Therefore, although liver and kidney both converted HMTBA to Met, it was retained by the former and exported by the latter. Under these experimental conditions, synthesis of Met from HMTBA completely eliminated use of dietary Met.

Comparative in vitro and in vivo absorption of 2-hydroxy-4(methylthio) butanoic acid and methionine in the broiler chicken.

Poultry diets are typically supplemented with DL-2-hydroxy-4(methylthio) butanoic acid (HMTBA, or the hydroxy analog of methionine) or DL-methionine (DLM). Although HMTBA and DLM provide methionine activity, they are structurally distinct molecules with different physiological characteristics until they are converted to L-methionine. The relative rates of intestinal HMTBA vs. DLM absorption have been controversial, and it has been claimed that HMTBA is not fully absorbed. We measured the uptake of HMTBA and DLM in an in vitro everted intestinal slice model. Sections of intestinal slices (jejunum and ileum) were incubated with 0.1 to 50 mM HMTBA that was radiolabeled or DLM that was radiolabeled, and absorption was measured by scintillation counting. The HMTBA uptake was equal to or greater than DLM absorption in each tissue and at every time point with one exception. Furthermore, the rates of HMTBA absorption were always equal to or significantly greater than DLM uptake. In a separate in vivo experiment, absorption of HMTBA and L-methionine was monitored along the entire gastrointestinal (GI) tract. Broilers were fed commercial-type corn-soy diets supplemented with 0.21% HMTBA. Digesta was collected from crop, proventriculus, gizzard, duodenum, jejunum, ileum, large intestine, and cloaca and analyzed for the concentration of free HMTBA and free methionine in each compartment. These studies demonstrated that HMTBA is absorbed completely and along the entire GI tract, especially the upper GI tract. Furthermore, there was a higher concentration of free L-methionine than HMTBA in the digesta from every segment distal to the gizzard.

Antibiotic growth promoters in agriculture: history and mode of action.

This report will review the history of antibiotic growth promoter (AGP) use in the animal industry, concerns about development of antimicrobial resistance, and response in the European Union and United States to these concerns. A brief description of the history of legislation regarding feed use of antimicrobials in Denmark and the experience of animal producers following the 1998 ban will serve to illustrate the consequences on animal performance and health of withdrawing the approval for this use. The biological basis for antibiotic effects on animal growth efficiency will consider effects on intestinal microbiota and effects on the host animal and will use the germ-free animal to illustrate effects of the conventional microflora. The probability that no single compound will replace all of the functions of antimicrobial growth promoters will be considered, and methods to consolidate and analyze the enlarging database will be discussed.

Impact of glutamine and Oasis hatchling supplement on growth performance, small intestinal morphology, and immune response of broilers vaccinated and challenged with Eimeria maxima.

Seven hundred and twenty hatchling broilers were allotted to 12 treatment groups. Groups 1 and 2 were fasted for 48 h posthatch; groups 3 and 4 were fasted for 48 h followed by ad libitum access to a 1% glutamine (Gln) diet; groups 5 and 6 had ad libitum access to a common diet; groups 7 and 8 had access to a 1% Gln diet posthatch; groups 9 and 10 were fed regular Oasis hatchling supplement; and groups 11 and 12 were fed Oasis sprayed with 1% Gln for the first 48 h posthatch. The birds in treatment groups 2, 4, 6, 8, 10, 11, and 12 were vaccinated with Eimeria maxima posthatch, and all birds were orally challenged with high dose E. maxima on d 22. During the first 2 wk, birds in group 7 had the highest gain and feed efficiency among treatments (P < 0.01). Compared with birds in the nonGln groups, birds in the Gln group had higher gain, feed efficiency, and livability (P < 0.05). Among the Fast (groups 1 to 4), Feed (groups 5 to 8), and Oasis (groups 9 to 12) groups, birds in the Feed groups had the highest gain during d 0 to 21 (P < 0.01). During d 22 to 28, birds in the Fast groups had the lowest BW and livability (P < 0.01), and the nonvaccinated birds had lower gain and feed efficiency relative to vaccinated birds (P < 0.01). Birds in the Feed and Oasis groups had higher villus height (VH) of mid small intestine than Fast groups at d 2 and 7 (P < 0.05), and nonvaccinated birds had higher VH than vaccinated birds (P < 0.01) at d 7 after hatch. On d 14, there were differences in serum interferon-gamma (P < 0.05) levels among treatments. During d 22 to 28, vaccinated birds had lower lesion scores in the mid small intestine than nonvaccinated birds (P < 0.01), and birds in the Feed or Oasis groups had lower lesion scores compared with the Fast groups (P < 0.02). These results indicated the importance of immediate access to feed posthatch, the beneficial effects of feeding Oasis hatching supplement and Gln after hatch, as well as the necessity of the vaccination program against coccidiosis challenge.

The effect of early posthatch starvation on calpain mRNA levels.

The calpain system is a family of calcium activated proteases that degrade myofibrillar protein. Male broiler chickens (Ross) were provided a standard starter diet top-dressed with Oasis((R)) nutritional supplement (fed; Novus International, St. Louis, MO, USA), or they were not provided any feed (starved) for the first 3 days posthatch. Subsequently, the standard starter diet was provided to all chickens between 3 and 7 days posthatch. RNA was extracted from the Pectoralis thoracicus, and skeletal muscle-specific n-calpain-1 (p94) calpain, mu-calpain, and m-calpain expression was evaluated using quantitative Northern analysis. Early posthatch starvation did not (P>0.05) affect calpain mRNA levels on each day examined. Similarly, there were no (P>0.05) changes in mu-calpain or m-calpain mRNA levels between 0 and 7 days posthatch in fed birds. However, p94 calpain mRNA levels were significantly (P<0.05) lower at 7 days posthatch compared to 0 or 2 days posthatch. Therefore, in the early posthatch chicken, it appears that the calpain system may not be affected by the presence of oral nutrition, and that there is an age-related downregulation of p94 calpain mRNA expression.

Uptake of DL-2-hydroxy-4-methylthio-butanoic acid (DL-HMB) in the broiler liver in vivo.

The methionine source DL-2-hydroxy-4methylthio-butanoic acid (DL-HMB; Alimet feed supplement) is widely used in the poultry industry. The purpose of this study was to determine the capacity of the broiler liver to remove DL-HMB from the circulation. Cannulae were implanted in the carotid artery and hepatic and hepatic portal veins in anesthetized male broilers (3.33 +/- 0.13 kg BW). In Experiment 1, birds (n = 5) were infused with DL-HMB solutions (diluted in saline, pH 7.2 to 7.4) into the hepatic portal vein at rates ranging from 4.4 to 22 mg/min per kg BW, whereas in Experiment 2, birds (n = 6) were infused with DL-HMB at rates ranging from 2.2 to 4.4 mg/min per kg BW. Plasma samples from each vessel were obtained before and after each 10-min DL-HMB infusion period with a 10-min clearance period allowed between each DL-HMB infusion. Regression analysis revealed a highly significant correlation in the amount of DL-HMB entering the liver via afferent vessels (afferent DL-HMB) and DL-HMB removed by the liver (y = 0.86(x) - 173, r2 = 0.98). The slope of this regression indicates that 86% of DL-HMB entering in afferent blood (i.e. from both the hepatic artery and hepatic portal vein) was removed or that the liver apparently metabolized 86% of the DL-HMB that entered the liver. The results indicate that the broiler liver has the capacity to remove DL-HMB from the circulation far in excess of that needed to metabolize DL-HMB that would enter the liver following gastrointestinal absorption in birds fed a conventional poultry diet. In addition, present results implicate the liver as a major site of removal from circulation and further metabolism of DL-HMB in chickens.

The hepatic extraction of plasma free amino acids and response to hepatic portal venous infusion of methionine sources in anesthetized SCWL males (Gallus domesticus).

This study was conducted to investigate the hepatic extraction of plasma free amino acids in anesthetized Single Comb White Leghorn (SCWL) males (Gallus domesticus). SCWL males were anesthetized and implanted with cannulae in the carotid artery, hepatic vein, hepatic portal vein and the left hepatic duct. Free amino acids in plasma and bile were determined before, during and after 30-min infusions of Saline (control), DL-Methionine (DL-Met) or DL-2-hydroxy-4-methylthio-butanoic acid (DL-HMB) into the hepatic portal vein. Hepatic extraction rates (HER) of amino acids were calculated based on the concentration of amino acids in plasma multiplied by estimations of blood flow in the hepatic portal vein, hepatic artery and hepatic vein. For the non-essential amino acids, alanine had the highest HER (46%). The liver also removed more than 20% of hepatic inflow of tyrosine and asparagine with substantial extraction (14-18%) of serine, glycine and glutamine, also. In contrast, less than 5% of hepatic inflow of glutamate and cystine were removed by liver. For the essential amino acids, HER for methionine, histidine and phenylalanine were 30, 14 and 17%, respectively, with less than 5% for branched-chain amino acids, lysine, arginine and threonine. Biliary secretion of amino acids represented a small percentage (<0.2%) of total hepatic extraction turnover of the amino acids. Infusion of methionine sources, DL-Met and DL-HMB, had no effect on hepatic metabolism of amino acids other than methionine. The results demonstrated for the first time, the hepatic extraction of circulating free amino acids in avian species in vivo.

Absorption of 2-hydroxy-4-(methylthio)butanoic acid by isolated sheep ruminal and omasal epithelia.

Alimet (Novus Inter., Inc., St. Louis, MO) feed supplement (an 88% aqueous solution of 2-hydroxy-4-(methylthio) butanoic acid; HMB) is a source of L-Met commonly used in nonruminants and ruminants. The absorption of HMB across ovine omasal and ruminal epithelia was evaluated in this study. Ruminal and omasal epithelia were collected from eight lambs (BW = 67.6 kg +/- 9.1) and mounted in parabiotic chambers that were repeatedly sampled throughout a 60-min incubation. The appearance of HMB (using DL-[5-14C]-HMB as a radiolabeled marker) in serosal buffers increased quadratically (P < .004) with time in both tissues. More (P < .001) HMB appeared in the serosal buffers with omasal than with ruminal epithelia. Both tissues responded similarly, and, after 60 min of incubation, the accumulation of HMB within the tissues increased linearly (P < .001) as substrate concentration (.375, .75, 1.5, 3.0, 6.0, and 12.0 mM) increased in mucosal buffers. As the concentration of HMB in the mucosal buffers increased, there was a quadratic (P < .001) increase in the appearance of HMB in the serosal buffer of the omasal epithelium, indicating some saturation of the system. The increase in serosal appearance of HMB was linear (P < .001) with ruminal tissue. The results indicate that there are probably multiple mechanisms involved in the absorption of HMB. Because saturation was observed in the omasum, it is likely that mediated transport accounts for at least a portion of the absorption of HMB in the omasum. Other mechanisms (e.g., diffusion and(or) paracellular absorption) are responsible for the balance of the absorption. Omasal epithelium appears to have a greater capacity for HMB absorption than ruminal epithelium. The enzymes involved in the conversion of HMB to 2-keto-4-(methylthio)butanoic acid were found in ruminal and omasal epithelia, liver and kidney. These results indicate that HMB can be absorbed across ruminal and omasal epithelium and that HMB can be used as a source of L-methionine.

Effect of turkey (Meleagridis gallopavo) breeder hen age and egg size on poult development. 2. Intestinal villus growth, enterocyte migration and proliferation of the turkey poult.

Villus growth, enterocyte migration and proliferation were measured in the small intestine of poults (Meleagridis gallopavo) to determine if hen age and/or egg size influences these characteristics during the first week after hatching. At hatching, distal jejunal villi were 22.8 microns longer in poults from the older (48 weeks) versus the younger (34 weeks) hens (P < 0.05). Similarly, labeled enterocytes in distal jejunal sections from poults from the older hens had migrated 28 microns (10%) farther along the crypt-villus axis at hatching, as compared to poults from the younger hens (P < 0.05). Villus growth differences and enterocyte migration were not consistently affected by hen age or egg weight class in poults from 1 to 7 days old. These results suggest that even though intestinal villi may be more advanced developmentally at hatch in poults from the older hens, however post-hatch growth of the intestine or the poult is not affected by hen age or egg weight class.

The relative effectiveness of 2-hydroxy-4-(methylthio) butanoic acid and DL-methionine in young swine.

We compared the effectiveness of 2-hydroxy-4-(methylthio) butanoic acid (HMB) and DL-methionine (DLM) as sources of L-methionine activity in methionine-deficient primary cultures of pig liver cells and methionine-deficient early-weaned pigs. Viable hepatocytes were obtained from minced pig liver and maintained in a high density, differentiated, nonproliferation cell culture system. Culture medium was supplemented with HMB, DLM, or L-methionine, and cells were pulse-dosed with L-[14C(U)]leucine for 24 h to determine the level of protein synthesis. Leucine incorporation per milligram of protein indicated a six-to eightfold increase in protein synthesis (P < .01) with methionine levels between 5 and 10 microM, regardless of source of methionine activity. Two 24-pen replicate methionine dose titrations were conducted with 95 early-weaned commercial crossbred pigs. The pelleted corn, dried whey, and porcine plasma basal diet contained 1.5% lysine, .23% methionine, and .48% cystine and was supplemented with 0, .05, or .10% methionine activity as DLM or HMB for 21 d. There was a 134, 104, and 61% increase (P < .01) in cumulative ADG for each successive week on study with a 30 and 19% improvement in feed/gain (P < .01) after 7 and 14 d. Performance responses due to source of methionine activity did not differ and slope ratio potency determinations (gain vs intake of methionine source) of HMB vs DLM indicated a 119, 111, and 95% relative activity for cumulative weekly performance. These results support the hypothesis that HMB and DLM provide equimolar levels of methionine activity in swine.

Optimum dietary arginine:lysine ratio for broiler chickens is altered during heat stress in association with changes in intestinal uptake and dietary sodium chloride.

1. The effects of varying the dietary arginine:lysine (Arg:Lys) ratio for broiler chickens at thermoneutral and high temperatures was studied in a series of 5 experiments which measured intestinal epithelial transport or evaluated growth and food efficiency with practical diets or diets supplemented with L-arginine free base. 2. The growth studies showed that increasing the Arg:Lys ratio at high temperatures produced consistent improvements in food conversion without any loss in growth. 3. Increasing dietary sodium chloride concentration reduced the Arg:Lys ratio necessary for optimum food conversion. 4. Food conversion responses were improved whether L-arginine free base was used as a dietary supplement in place of an inert filler or practical diets with differing ingredients were used to vary the Arg:Lys ratio. 5. In the presence of an equimolar concentration of lysine the uptake of arginine by the intestinal epithelium of heat-stressed broilers was reduced significantly compared with that of broilers at thermoneutral temperatures. 6. The results indicate that the ideal amino acid balance for broilers varies with ambient temperature.

Effect of intermittent periods of high environmental temperature on broiler performance responses to sources of methionine activity.

The objective of these experiments was to study the effect of intermittent high temperature (IHT) on performance of broilers fed different sources of methionine activity. Two 20-d experiments were conducted in which individually caged 28-d-old cockerels were exposed to a 5-d period of constant high temperature (HT, 30 to 32 C) followed by an equal period of thermoneutral (TN) temperatures (22 C) with the 10-d temperature cycle repeated twice. Birds held at 22 C were TN controls. Grower diets contained 3,275 kcal/kg ME and 20% CP and were supplemented with either 2-hydroxy-4-(methylthio)butanoic acid (HMB, Alimet Feed Supplement) or DL-Met up to a maximum of .88 to .90% total sulfur amino acids (TSAA). In Experiment 1, gain:feed but not average daily gain was greater (P < .05) for HMB than DL-Met birds subjected to IHT (.451 vs .413, respectively), but no treatment differences were observed for TN birds. Results of Experiment 2 demonstrated a linear response to HMB and DL-Met dose in TN (P < .01); however, only HMB-supplemented birds responded similarly in IHT (P < .01). These results are consistent with lower availability of DL-Met as a result of IHT. In vitro experiments indicated that capacity for uptake of DL-Met into intestinal epithelial cells was reduced in heat-stressed birds. Uptake of D-Met was more severely affected than was L-Met. Consequently, a third performance experiment compared the ability of D-Met and L-Met to support growth under conditions of HT. The results indicated that the effect of HT on broiler performance was mediated through reduced utilization of D-Met.

Responses of laying hens to diets containing up to 2% DL-methionine or equimolar (2.25%) 2-hydroxy-4-(methylthio)butanoic acid.

Diets supplemented with up to .6% DL-Met (DLM) or .68% 2-hydroxy-4-(methylthio)butanoic acid (HMB, Alimet) acidify the urine and reduce the incidence of urolithiasis in pullets and laying hens. Excessive acidification potentially may reduce eggshell quality and bone mineralization by interfering with Ca metabolism and may severely challenge the liver and kidneys, which are the primary organs responsible for attenuating metabolic acidosis. To evaluate these possibilities, 30-wk-old Single Comb White Leghorn hens in full production (five hens per replicate, six replicates per diet treatment) were fed for 30 d a 15.7% CP corn and soybean meal-based control layer ration alone or supplemented with DLM (.5, 1, 1.5, or 2%) or equimolar HMB (.56, 1.13, 1.69, or 2.25%). None of the diets caused mortality or gross hepatic or renal damage. Hens fed diets supplemented with the highest levels of DLM and HMB exhibited significant reductions in feed intake, hen-day egg production, and liver mass and had lower plasma concentrations of alanine amino-transferase and isocitrate dehydrogenase when compared with hens fed the control diet. Kidney mass was not significantly affected by high levels of DLM or HMB, but plasma uric acid was significantly higher in hens fed 2% DLM compared with hens fed the control diet. The highest levels of DLM and HMB did not significantly alter total plasma Ca or inorganic phosphate concentrations, nor were percentage eggshell or femur mineralization (femur ash mass:defatted bone mass, femur ash mass:bone volume) significantly reduced.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical and nucleic acid analysis of somatotropin receptor populations in the bovine ovary.

Ovaries were analyzed for somatotropin receptor protein and mRNA through use of immunohistochemistry, solution hybridization/nuclease protection, Northern blotting, and reverse transcriptase polymerase chain reaction (RT-PCR). As indicated by immunoperoxidase staining, CL expressed immunoreactive somatotropin receptor (positive stain). Ovarian stroma, connective tissue, endothelium, and erythrocytes did not express somatotropin receptor (negative stain). Within the CL, somatotropin receptor protein was expressed primarily in large luteal cells whereas small luteal cells were negative. Most follicles (1-5 mm, after fixation) were negative for somatotropin receptor. On the basis of solution hybridization/nuclease protection, the mRNA for somatotropin receptor was found in greatest abundance in CL and large luteal cells and was nearly undetectable in small luteal cells or follicles (class 1, 3-5 mm; class 2, 6-9 mm; and class 3, > or = 10 mm). Northern blotting of mRNA for somatotropin receptor showed expression of somatotropin receptor mRNA transcripts in whole ovary (4.7 and 4.4 kb), CL (4.7 and 4.4 kb), and liver (4.4 kb); and RT-PCR amplified a single amino acid coding region for somatotropin receptor in CL and liver. In summary, somatotropin receptor (both immunoreactive protein and mRNA) is found primarily in the large luteal cell, and lesser amounts of the expressed receptor or its message are found in the follicle. Alternative sizes of mRNA for somatotropin receptor suggest novel mRNA processing in the bovine ovary.

Effect of heat stress on 2-hydroxy-4-(methylthio)butanoic acid and DL-methionine absorption measured in vitro.

The objective of the present experiments was to determine the biochemical basis for preliminary chick performance data, which indicate an ameliorative effect of 2-hydroxy-4-(methylthio)butanoic acid (HMB) when compared with DL-methionine (DLM) fed during hot conditions. In vitro passage of HMB or DLM across intact segments of small intestine from either control (thermoneutral, TN) or heat-stressed (HS) birds was used as a model for intestinal absorption. For DLM placed in the lumen, appearance in the outside buffer was reduced when using intestine from HS birds compared with tissue from TN birds. In contrast, the appearance of HMB in the outside buffer was greater using HS intestine, resulting in a substrate by environment interaction (P < .01). Slices of everted small intestine from TN and HS birds were used to study epithelial uptake of methyl labeled 14C-DLM by three transport pathways: diffusion, carrier-specific energy- and sodium-independent uptake (ESI), and carrier-specific energy- and sodium-dependent uptake (ESD). Correcting for extracellular volume, total epithelial uptake of 14C-DLM (diffusion plus ESI plus ESD) was reduced by 34% in HS intestine (P < .05). Energy-dependent uptake was observed to decrease by 87% in HS (P < .05). Energy-independent uptake was increased (136%, HS versus TN, P < .05), but not enough to compensate for the decrease in ESD uptake. Intestinal transport systems for glucose and leucine were also observed to change during HS, suggesting a role for cellular transport changes in the performance reduction associated with HS.