Determination of TMEn, standardized amino acid digestibility, phosphorus digestibility, and phosphorus bioavailability in conventional and reduced phosphorus distillers dried grains with solubles.

The TMEn, amino acid (AA) digestibility, and P availability in 2 conventional corn distillers dried grains with solubles (C-DDGS1 and 2; 0.86 to 1.14% P, DM basis) and reduced phosphorus DDGS (RP-DDGS; 0.39% P) were evaluated. The TMEn of C-DDGS1 and 2 and RP-DDGS were determined in Experiment 1 using conventional adult Leghorn roosters, while standardized AA digestibility was determined in Experiment 2 using cecectomized roosters. Apparent ileal digestibility (AID) of P at different Ca levels was determined using precision-fed (crop intubation) broiler chickens in Experiments 3 and 4. The AID and total tract retention of P in C-DDGS2 were evaluated in Experiment 5 using ad libitum-fed broilers. Phosphorus bioavailability in C-DDGS2 relative to KH2PO4 based on bone ash was determined in Experiment 6. Experiments contained 4 to 5 replicates per treatment. In Experiment 1, the TMEn of C-DDGS1 and RP-DDGS was 3,428 and 2,840 kcal/kg, respectively (DM basis). In Experiment 2, there were no differences (P > 0.05) in rooster AA digestibility values between C-DDGS1 and RP-DDGS. In Experiment 3 with precision-fed chicks, AID of P in C-DDGS1 and RP-DDGS was 81 and 59%, respectively; there was no effect (P > 0.05) of increasing dietary Ca level from 0.04 to 1.0% for C-DDGS1 or reducing Ca from 1.5 to 1.0% for RP-DDGS. The AID of P in precision-fed chicks for C-DDGS2 in Experiment 4 was 48 and 80% at 1.3 and 0.3 Ca:total P ratios, respectively (P < 0.05). In Experiment 5, AID of P in C-DDGS2 at Ca:total P ratios of 1.3 and 2.5 was 63 and 42%, respectively, in precision-fed chicks. Regression of bone ash content (mg/tibia) on supplemental P intake in Experiment 6 yielded a P bioavailability of 61% relative to KH2PO4 for C-DDGS2. In conclusion, total and digestible P content in RP-DDGS was greatly reduced compared with C-DDGS, and the digestibility and bioavailability of the P in C-DDGS was affected by type of experimental assay and dietary Ca level.

Evaluation of the efficacy of vitamin D3 or its metabolites on thiram-induced tibial dyschondroplasia in chickens.

Two trials were conducted to determine if thiram-induced tibial dyschondroplasia (TD) in chickens was linked to a vitamin D deficiency and calcium homeostasis dysregulation, and whether feeding vitamin D fortified diets may prevent it. Day-old chickens were given grower diets containing different vitamin D products throughout the experiment until necropsy on day 16. Half of the birds in each feed group received thiram at levels of 100 ppm (trial 1) or 50 ppm (trial 2) between days 7-9 to induce TD. The birds were weighed, bled, and euthanized to determine TD incidences and severity by examining the growth plates. Tibial bones were used to measure biomechanical strength and ash content. Blood concentrations of 25-hydroxyvitamin D, Ca, P, alkaline phosphatase, and creatine kinase were measured in serum that showed no differences between different groups. Thiram reduced body weight and induced TD regardless of any vitamin D treatment to the same extent as untreated birds.

Efficacy of an Escherichia coli phytase in broilers fed adequate or reduced phosphorus diets and its effect on carcass characteristics.

Five experiments (EXP) were conducted to assess the efficacy of an Escherichia coli phytase compared with 2 commercially available fungal phytases. In EXP 1 and 2, male broiler chicks were fed experimental diets that included a P-deficient control (0.13% available P; 0.88% Ca) alone or with graded levels of KH(2)PO(4) (0, 0.05, 0.10, or 0.15%) or phytase at levels of 250, 500, 1,000, 2,000, or 4,000 phytase units/kg of E. coli phytase (EXP 1 and 2), fungal phytase 1 (EXP 2), or fungal phytase 2 (EXP 2). In EXP 1 and 2, weight gain and tibia ash (mg/chick and %) responded linearly (P < 0.05) to inorganic P addition. In EXP 2, each level of E. coli phytase released more P than either fungal phytases 1 or 2, whether based on tibia ash weight (mg/chick) or percentage. In EXP 3, 4, and 5, dietary treatments containing adequate or deficient levels of P were fed with or without supplemental E. coli phytase. In EXP 3, weight gain and tibia ash were reduced (P < 0.05) by P deficiency, but gain and tibia ash of chicks fed E. coli phytase (250, 500, or 1,000 phytase units/kg) did not differ (P > 0.05) from that of chicks fed the P-adequate diet. In addition, carcass yield of broilers fed E. coli phytase was not reduced (P > 0.05). In EXP 4, E. coli phytase effectively supported weight gain, tibia ash, breast yield, and leg yield compared with birds fed the P-adequate diet, but clavicle breakage during processing was increased in birds fed E. coli phytase. In EXP 5, E. coli phytase again effectively supported weight gain, and no differences (P > 0.05; compared with the P-adequate diet) were noted for clavicle ash, diameter, or breaking strength. No differences (P > 0.05) in bone breakage during processing were noted among treatments. These results indicate that the addition of E. coli phytase to P-deficient broiler diets improves growth, bone, and carcass performance and is more effective at releasing phytate-bound P than the other phytase products that were tested.

Effect of reciprocating dietary lysine fluctuations on chick growth and carcass yield.

Three experiments (Exp) were conducted to study the effect of reciprocating (i.e., back-and-forth) fluctuations in dietary Lys concentration on growth performance of male broiler chicks. The Lys, CP, and ME concentrations were fluctuated in Exp 1 by varying corn and soybean meal concentrations. Corn-peanut meal diets in Exp 2 and 3 contained a constant calorie-protein ratio, but Lys concentrations were fluctuated by varying supplemental L-Lys x HCl. During 7-d feeding periods from d 8 to 43 posthatch, chicks fed the fluctuating regimen were provided diets with nutrient concentrations arranged in the following order (relative to control diets at any given point): excess-deficient-excess-deficient-adequate. In each Exp, differences in feed intake and gain:feed were observed during the first 7 d. Weight gain was never affected by dietary regimen. Overall, chicks fed the fluctuating regimen showed similar weight gain and feed intakes relative to chicks fed the control regimen. Additionally, efficiency of feed, Lys, CP, and ME utilization over the 35-d growth period of Exp 2 and 3 were not affected by dietary regimen. An increase in carcass yield (dressed, carcass weight as a percentage of live weight) of birds fed the fluctuating regimen was observed, but there were no differences in abdominal fat yield (as a percentage of dressed, carcass weight). These data support the conclusion that reciprocating 7-d periods of nutrient excess and deficiency do not affect overall growth performance of growing chicks.

Homocysteine remethylation in young broilers fed varying levels of methionine, choline, and betaine.

Methionine is critical in amino acid nutrition for chickens, yet details of the flux of Met metabolites in the avian system are lacking. This study explored the interactions among dietary choline (CHO), betaine (BET), and sulfur amino acid levels on growth and hepatic homocysteine (HCY) remethylation. Graded levels (0, 0.07, 0.11, and 0.24%) of DL-Met were added to diets adequate in CHO and deficient in sulfur amino acids (0.26% digestible Met, 0.26% digestible Cys). Each Met level was tested alone or with the addition of CHO (0.25%) or BET (0.28%). Broilers were reared from 8 to 22 d in raised wire floor battery cages, and the 12 dietary treatments were fed to 3 replicate pens containing 5 birds per pen. Weight gain and feed efficiency were maximized (P < 0.05) with addition of 0.11% supplemental Met, whereas feed intake was maximized (P < 0.05) with addition of 0.07% supplemental Met. Overall, growth parameters were not affected (P > 0.05) by CHO or BET addition. Hepatic tissue primed by the different dietary treatments was subjected to a newly developed stable isotope methodology and HPLC-mass spectrometry to quantify the impact of diet on HCY remethylation. Dietary Met level did not (P > 0.05) affect HCY remethylation, but remethylation through the Met synthase pathway was increased (P < 0.05) by addition of CHO or BET to diets containing deficient or excess levels of Met. Minimal changes in hepatic HCY remethylation through the betaine-homocysteine methyltransferase pathway occurred in response to dietary changes; therefore, data failed to support previous suggestions that BHMT might have a regulatory role when diets containing deficient or excess Met levels are fed. In contrast to previous suppositions based on enzyme activity, under most dietary conditions, the quantity of HCY remethylated by Met synthase appeared to exceed that remethylated by the alternate betaine-homocysteine methyltransferase pathway.

Protein and body weight accretion of chicks on diets with widely varying contents of soyabean meal supplemented or unsupplemented with its limiting amino acids.

1. Two studies were conducted to evaluate the effects of relative deficiencies of methionine and threonine on the growth performance of 8- to 21-d-old chicks fed on isoenergetic diets containing a wide range of crude protein (CP) concentrations from dehulled soyabean meal (SBM). 2. Chicks fed on graded levels of SBM containing supplemental methionine and threonine (BAL) accreted whole-body protein more efficiently (P < 0.05) than those receiving graded levels of unsupplemented SBM (DEF), and superior (P < 0.05) growth performance was also obtained at lower CP levels when chicks were fed on the BAL diets. 3. Voluntary food intake increased between 30 and 220 g CP/kg in chicks fed on DEF diets, whereas food intake of chicks fed on BAL diets increased only between 30 and 100 g CP/kg, after which it decreased between 100 and 220 g CP/kg. 4. Protein efficiency ratio (g gain per g protein intake) decreased with each incremental increase in CP between 30 and 260 g CP/kg, regardless of whether diets were BAL or DEE 5. These data indicate that maintaining a balanced ratio of amino acids is a preferable approach when poultry producers are interested in employing low CP diets for economic, physiological or environmental reasons.

Single versus multiple deficiencies of methionine, zinc, riboflavin, vitamin B-6 and choline elicit surprising growth responses in young chicks.

A soy-protein isolate diet that was deficient in methionine (Met), zinc (Zn), riboflavin, vitamin B-6 and choline for chick growth (Assay 1) was used to study individual or multiple deficiencies of several of these nutrients. In all cases, adding all three deficient nutrients together resulted in growth responses that were superior to those resulting from supplementation with any pairs of deficient nutrients. In Assay 2, single addition of Zn but not of methionine or riboflavin produced a growth response, but the combination of either Zn and Met or Zn and riboflavin resulted in growth responses that were greater than the response elicited by Zn alone. Assay 3 involved individual or multiple deficiencies of choline, riboflavin and vitamin B-6, and individual additions suggested that choline was first limiting. Choline + riboflavin supplementation, however, produced marked growth and gain:food responses that were far greater than those resulting from supplemental choline or riboflavin alone. Moreover, the growth response to a combination of choline + pyridoxine (PN) was also greater than that obtained from any of the three nutrients fed alone; even PN + riboflavin (in the absence of choline) produced responses greater than those observed with the unsupplemented negative-control diet. In Assay 4, chicks responded to individual additions of riboflavin, PN or Met, and in Assay 5, to either riboflavin or PN; all two-way combinations resulted in growth rates that were far greater than those occurring with any single addition. The data from these experiments show that unlike the situation with three deficient amino acids, the expected responses to first-, second- and third-limiting B-vitamins or deficient vitamins combined with deficient levels of Zn or Met do not follow the expected pattern of response to first-, further response to first- and second- and an even further response to first-, second- and third-limiting nutrients.

Bioavailability of lysine from a liquid lysine source in chicks.

Two experiments were conducted to assess the bioavailability of lysine from a liquid lysine product (LLP; 60% lysine) relative to crystalline L-lysine.HCl. In the first experiment, four groups of five chicks were fed a lysine-deficient basal diet containing corn, soybean meal, and feather meal or the basal diet supplemented with 0.1 or 0.2% lysine from L-lysine.HCl or LLP from Day 8 to 22 posthatching. Weight gain and feed efficiency responded linearly (P < 0.01) to increasing levels of lysine from either lysine source, and multiple linear regression analysis of weight gain regressed on supplemental lysine intake indicated no difference (P > 0.05) in the response to lysine from L-lysine.HCl or LLP. Experiment 2 was conducted using a lysine-deficient basal diet containing corn, peanut meal, and feather meal, and all diets were fed to five groups of five chicks from Day 8 to 21 posthatching. Treatment additions again consisted of 0.1 or 0.2% lysine from L-lysine.HCl or LLP. Weight gain and feed efficiency responded linearly (P < 0.01) to increasing levels of lysine from L-lysine-HCl and LLP, and multiple linear regression analysis of weight gain regressed on supplemental lysine intake indicated no difference (P > 0.05) in the response to lysine from L-lysine.HCl or LLP. These data indicate that lysine from LLP is fully bioavailable relative to lysine from crystalline L-lysine.HCl, and could therefore be used as a source of lysine in practical poultry diets.

Limiting order of amino acids in a low-protein corn-soybean meal-whey-based diet for nursery pigs.

Three trials were carried out with pigs between 5 and 8 wk of age to determine the limiting order of amino acids in a 13.5% CP corn-soybean meal-based diet containing 8% dried whey. The positive-control diet was a 19.2% CP corn-soybean meal-based diet (1.15% lysine), also with 8% dried whey. Amino acid additions to the low-protein, negative-control diet were based on levels needed to accomplish 110% of ideal ratios (to lysine, set at 1.15%). In Exp. 1, the addition of an amino acid mixture containing Lys, Trp, Thr, Met, Ile, and Val to the low-protein diet increased (P<.05) gain and gain: feed ratio, and these response traits were not different from those of pigs fed the 19.2% CP positive-control diet. Single deletion of Lys from the supplemental amino acid mixture depressed performance to a greater (P<.05) extent than single deletion of any of the other amino acids. Single deletions of Trp, Thr, Met, or Val decreased (P<.05) performance in a similar but lesser magnitude than the decrease caused by Lys deletion, whereas Ile deletion was without effect. Experiments 2 and 3 were designed to evaluate the limiting order of AA beyond Lys in the low-protein diet. Neither His nor Glu were found to be deficient, and, as in Exp. 1, deletion of Trp, Thr, Met, or Val from the supplemental amino acid mixture resulted in performance depressions (P<.05) that were similar. The results suggest that Lys is first-limiting and Trp, Thr, Met, and Val are equally second-limiting in a reduced protein (13.5% CP) corn-soybean meal-based diet with 8% whey for 10-kg pigs.

Bioavailability of zinc in two zinc sulfate by-products of the galvanizing industry.

Two Zn depletion/repletion assays were conducted with chicks to determine the relative bioavailability (RBV) of Zn from two new by-products of the galvanizing industry. Using a soy concentrate-dextrose diet, slope-ratio methodology was employed to evaluate two different products: Fe-ZnSO4 x H2O with 20.2% Fe and 13.0% Zn, and Zn-FeSO4 x H2O with 14.2% Fe and 20.2% Zn. Feed-grade ZnSO4 x H2O was used as a standard. Weight gain, tibia Zn concentration, and total tibia Zn responded linearly (P < 0.01) to Zn supplementation from all three sources. Slope-ratio calculations based on weight gain established average Zn RBV values of 98% for Fe-ZnSO4 x H2O and 102% for Zn-FeSO4 x H2O, and these values were not different (P > 0.10) from the ZnSO4 standard (100%). Slope-ratio calculations based on total tibia Zn established average Zn RBV values of 126% for Fe-ZnSO4 x H2O and 127% for Zn-FeSO4 x H2O, and these values were greater (P < 0.01) than those of the ZnSO4 standard (100%). It is apparent that both mixed sulfate products of Fe and Zn are excellent sources of bioavailable Zn.

Bioavailability of iron in cottonseed meal, ferric sulfate, and two ferrous sulfate by-products of the galvanizing industry.

Iron depletion-repletion assays were carried out with young chicks to establish Fe bioavailability values for Fe2(SO4)3.7H2O (22.7% Fe), Fe-ZnSO4.H2O (20.2% Fe, 13.0% Zn), Zn-FeSO4.H2O (20.2% Zn, 14.2% Fe), and cottonseed meal (200 mg Fe/kg). Standard hemoglobin response curves were established using feed-grade FeSO4.H2O (28.8% Fe) or reagent-grade FeSO4.7H2O (20.1% Fe) as standards such that relative bioavailability (RBV) could be assessed for the experimental sources of Fe. Weight gain, hemoglobin, and hematocrit responded linearly (P < 0.05) to Fe supplementation in all assays. Using hemoglobin as the response criterion, slope-ratio calculations established Fe RBV values of 126% for Fe-ZnSO4.H2O and 93% for Zn-FeSO4.H2O. The 126% value for Fe-ZnSO4.H2O was greater (P < 0.05) than the FeSO4.H2O standard (100%), but the 93% value for Zn-FeSO4.H2O was not different (P > 0.10) from the standard. However, evaluation of all criteria of response (hemoglobin, hematocrit, weight gain) suggested that neither Fe-ZnSO4.H2O nor Zn-FeSO4.H2O had different Fe RBV values than FeSO4.H2O. Standard-curve calculations were used for assessment of Fe RBV in Fe2(SO4)3.7H2O and cottonseed meal, as only a single level of Fe addition was studied for each of these products. Iron RBV in Fe2(SO4)3.7H2O was estimated to be 37%, whereas Fe RBV in cottonseed meal was found to be 56%. Both of these values were lower (P < 0.05) than the FeSO4 standard. The data suggest that the two new products, representing combinations of FeSO4.H2O and ZnSO4.H2O by-products of the galvanizing industry, are excellent sources of bioavailable Fe, whereas ferric sulfate and cottonseed meal are relatively poor sources of usable Fe.

Vitamin D3 requirement of young chicks receiving diets varying in calcium and available phosphorus.

1. Three battery experiments were conducted with broiler chicks during the 2nd and 3rd week of life. Graded amounts of cholecalciferol (D3) were added to maize-soyabean meal diets that were designed to be (a) severely deficient in available phosphorus (P), (b) marginally deficient in calcium (Ca) or (c) adequate in both available P and Ca. 2. With diets containing 1.0 g available P and 6.3 g Ca/kg (assay 1), graded doses of D3 between 0 and 37.5 mu/kg produced linear (P < 0.05) positive responses in both weight gain and tibia ash. With a D3 concentration of 1250 micrograms/kg, 250 times the requirement recommended by the NRC, bone ash was increased (P < 0.05) over that of birds fed 37.5 micrograms/kg, and neither weight gain nor food intake were reduced. 3. With a P-adequate diet (4.5 g available P/kg) containing 8.5 g Ca/kg (assay 2), weight gain and bone ash increased linearly (P < 0.05) upon supplementing the basal diet with 0, 2.5 and 5.0 micrograms D3/kg. Higher doses of D3 did not elicit further responses, and chicks fed on a diet containing 1250 micrograms D3/kg gained as fast and had bone ash values that did not differ from those of chicks receiving 5, 10, 20 or 40 micrograms D3/kg. 4. When the maize-soyabean meal basal diet was fortified with Ca and P to achieve adequate amounts of Ca (10.1 g/kg) and P (4.5 g available P/kg) in assay 3, dietary additions produced results similar to those obtained in assay 2 where P was adequate and Ca was slightly deficient. Again, chicks receiving a surfeit of D3 (1250 micrograms/kg) exhibited weight gains and bone ash values that were as great as those of chicks receiving 5, 10, 15 or 30 micrograms D3/kg. 5. It is apparent that young chicks have a high tolerance for excess D3, and chicks fed on diets that are severely deficient in available P continue to respond to D3 in excess of 37.5 micrograms/kg.

Hepatic and renal betaine-homocysteine methyltransferase activity in pigs as affected by dietary intakes of sulfur amino acids, choline, and betaine.

In Exp. 1, young pigs were fed a basal diet containing .17% methionine (Met) (.14% digestible Met), and .48% cystine (.38% digestible cystine) for 14 d (34 to 48 d of age). Treatment additions were .25% DL-Met, .34% betaine, .30% choline, or .25% DL-Met and .34% betaine. Methionine, but not betaine or choline supplementation, increased (P < .05) weight gain and feed efficiency. Hepatic betaine-homocysteine methyltransferase (BHMT) activity was increased (P < .05) by betaine and choline supplementation but was not affected by Met deficiency. Renal BHMT activity was increased (P < .05) by Met deficiency and was further increased (P < .05) by betaine supplementation. In Exp. 2, 10-kg pigs were fed the basal diet from Exp. 1 supplemented with enough DL-Met to bring the total basal Met to .24% (.20% digestible Met). Treatment additions consisted of .20% DL-Met or .34% betaine, and diets were fed for 16 d (34 to 50 d of age). Feed efficiency increased (P < .05) in response to Met, but not to betaine, supplementation. Hepatic BHMT activity increased (P < .05) in response to betaine and Met, but no changes in renal BHMT activity occurred. Although statistically significant changes in hepatic and renal BHMT activity occurred in both experiments, the magnitude of the responses was probably not physiologically important. Therefore, in contrast to previous findings with rats and chicks, it does not seem that hepatic and renal BHMT activity in pigs is influenced substantially by Met deficiency, or by surfeit levels of choline or betaine.

Iron bioavailability in soybean meal as affected by supplemental phytase and 1 alpha-hydroxycholecalciferol.

An Fe depletion-repletion chick bioassay was conducted to determine whether supplemental microbial phytase or 1 alpha-hydroxycholecalciferol (1 alpha-OH D3) would improve the bioavailability of Fe in soybean meal (SBM). Weight gain, hemoglobin, and hematocrit were markedly improved when increasing levels (0, 10, 20, and 80 mg/kg) of Fe from analytical grade ferrous sulfate (FeSO4.7H2O) were added to the Fe-deficient casein-dextrose basal diet containing 20 mg Fe/kg. Addition of 19 mg Fe/kg from SBM to the basal diet improved (P < 0.05) hemoglobin and hematocrit, but the response was less than that obtained from 10 mg Fe/kg from FeSO4.7H2O. Phytase (1,430 units/kg), 1 alpha-OHD3 (10 micrograms/kg), or the combination, added to the SBM-fortified basal diet did not further improve hematocrit or hemoglobin, indicating that Fe bioavailability of SBM was not increased by either of these feed additives. Based on standard-curve methodology, and using hemoglobin as a criterion, the relative bioavailability of Fe was 38.5% for SBM, 21.0% for SBM+phytase, 23.2% for SBM+1 alpha-OHD3, and 29.2% for SBM+phytase+ 1 alpha-OHD3.

Development of an experimental diet for determining bioavailable choline concentration and its application in studies with soybean lecithin.

Attempts to determine choline bioavailability have encountered criticism of experimental diets and protocol. Our objectives were to develop a choline-deficient soy isolate diet to quantify bioavailable choline concentration of soybean lecithin and to compare results to those obtained with a purified diet. In Assay 1, weight gain of chicks fed a choline-free crystalline amino acid diet responded linearly (P < .01) to graded doses of choline chloride, fluid lecithin (FL), or deoiled lecithin (DL). Multiple linear regression analysis indicated a bioavailable choline content of 2.3 and 3.7% for FL and DL, respectively. In Assay 2, a choline-deficient soy isolate diet was supplemented with 2-amino-2-methyl-1-propanol (AMP) to inhibit choline biosynthesis. Weight gain, feed intake, and feed efficiency increased (P < .05) markedly with the addition of choline chloride, but supplemental methionine or betaine had no effect (P > .10). Addition of 10% soybean meal to the diet severely deficient in choline per se produced a growth response (P < .05), whereas the same addition to the diet made adequate in choline did not elicit a growth response. In Assay 3, addition of graded levels of choline chloride, FL, or DL to the choline-deficient soy isolate diet containing AMP resulted in a linear (P < .01) increase in weight gain and feed intake. Multiple linear regression analysis indicated a bioavailable choline content of 2.0 and 3.5% for FL and DL, respectively. The AMP-containing soy isolate diet seems well suited for determination of bioavailable choline content in products containing secondary nutrients. Bioavailable choline content of FL and DL was similar to estimates of total choline content, suggesting that the choline in these products was fully available.

Hepatic betaine-homocysteine methyltransferase activity in the chicken is influenced by dietary intake of sulfur amino acids, choline and betaine.

There is much interest in the metabolism of homocysteine, because elevated plasma homocysteine [hyperhomocyst(e)inemia] is an independent risk factor for the development of cardiovascular disease. Four chick assays were conducted to determine the effects of varying dietary sulfur amino acids, choline and betaine on the activity of hepatic betaine-homocysteine methyltransferase (BHMT), an enzyme likely to be important in modulating plasma homocysteine. In Experiment 1, chicks were fed a purified crystalline amino acid diet containing adequate sulfur amino acids and choline. Excess dietary methionine, or the combination of excess cystine with choline or betaine, caused a small increase (P < 0.05) in BHMT activity. In Experiment 2, use of a methionine-deficient purified diet resulted in a threefold increase (P < 0.05) in BHMT activity, and addition of choline or betaine further increased (P < 0.05) BHMT activity. In Experiment 3, use of a methionine-deficient corn-peanut meal diet increased BHMT (P < 0.05) relative to that of chicks supplemented with adequate methionine, and addition of surfeit choline to the methionine-deficient basal diet caused a further increase (P < 0.05). In Experiment 4, addition of both surfeit choline and surfeit betaine to the methionine-deficient corn-peanut meal diet caused an increase (P < 0.05) in BHMT activity relative to that observed in chicks fed the methionine-deficient basal diet. These assays show that large increases in BHMT activity can be produced under methionine-deficient conditions, especially in the presence of excess choline or betaine.