Amino Acid Nutrition of the Young Calf. Estimation of Methionine and Lysine Requirements

Amino Acid Nutrition of the Young Calf. Estimation of Methionine and Lysine Requirements 1,2

D I A N A TZENG and C. L. D A V I S Department of Dairy Science

University of Illinois Urbana 61801

ABSTRACT

A semipurified diet containing 14 crystalline amino acids as the sole source of nitrogen was used qualitatively and quantitatively to determine dietary needs of the young calf for lysine and methionine.

Body weight gain, nitrogen balance, and concentrations of free amino acids in plasma were the criteria to assess the response of young calves to the experimental diets containing graded amounts of lysine and methionine. By these methods the methionine requirement as D-L methionine in the absence of cystine ranged from .17 to .23 g/day/kg body weight (.65 g/kg weight 'Ts ).

I N T R O D U C T I O N

Nitrogen nutrit ion of the young calf has been researched extensively in amount and source of protein in the diet (8, 10, 13, 14). However, few studies have been directed toward establishing amino acid requirements of the suckling calf. It generally has been assumed that the young suckling calf is as dependent upon certain amino acids in its diet as are nonruminant species (4) and that whole milk proteins supply an opt imum balance of amino acids for growth. The objectives of this study were 1) to develop a suitable semipurified diet containing crystalline amino acids as the sole source of nitrogen to be used as a test diet in future studies; 2) to ascertain essentiality of

Received August 16, 1979. t Part of thesis of senior author submitted to grad-

uate college in partial fulfillment of the requirement for Ph.D. in nutritional sciences at University of Illi- nois, Urbana-Champaign, 1974.

~This research was supported in part by Illinois Agricultural Experiment Station Project 35-335 and USPHS Predoctoral Training Grant GM00653.

methionine and lysine for the young calf; and 3) to establish quantitative requirements for these two amino acids by the young calf. Body weight change, nitrogen balance, and free amino acid profiles of plasma were the criteria for assessing responses to the diets.

M A T E R I A L S AND METHODS

Experimental Animals and Treatments

Twelve male calves (Holstein, Guernsey, and Jersey breeds) ranging in age from 3 to 60 days were in this study. Calves were housed in individual units at the Nutrit ion Laboratory of the Department of Dairy Science, University of Illinois.

In Experiment 1, three calves were in a switch-over design to evaluate the experimental diet. Each calf was subjected to four experimental periods. During periods 1 and 3 calves received the control diet which consisted of whole cow's milk (Table 1) fed at 10% of body weight. During periods 2 and 4 calves received the experimental diet at 10% of body weight. Between periods 1 and 2 calves were allowed 10 days to adjust to the experimental diet, whereas between periods 2 and 3 and 3 and 4 less t ime (3 days) was needed to assure desired consump- tion of the diet and to avoid digestive distur- bances. Once the calves had adjusted to the diet, the experimental phase was initiated in which total feed intake and body weight were measured, and total fecal and urinary outputs were collected for 5 consecutive days. Calves started on control diet (whole milk) at 3 days of age and proceeded through the t reatment periods ending up at 50 to 60 days of age.

In Experiments 2 and 3, nine calves were used to study qualitative and quantitative dietary requirements for lysine and methionine. During Experiment 2, calves received the same experimental diet as in Experiment 1 except that the lysine content of the diet was varied. The amounts of L-lysine-HC1 in the diet, as

1980 J Dairy Sci 63:441-450 441

442 TZENG AND DAVIS

TABLE 1. Chemical composition of diets.

Diets Cow's Experi-

Ingredients milk mental

(g/kg DM) Protein a 272 0 Amino acid mixb 0 339.2 Fat c 280 253.0 Lactose 371 335.0 Minerals d 55.8 75 Vitamins e 5.9 Sodium bicarbonate 0 3.6 Lecithin 0 7.5

aNitrogen × 6.38. b Composxtion of amino acid mixture (g/kg DM):

L-Arg -He1 10.15; L-HIS -HC1-H20 8.75; L-lsoleu 14.15; L-Leu 23.27; L-Lys -He1 24.27; DL-Met 6.24; L-Oala 11.75; L-Thr 11.04; L-Try 3.36; L-Val 15.84; L-Prol 23.75; L-Try 12.24; Gly 4.8; L-Glutamic acid 169.63.

CFat in experimental diet was lard. dMineral mixture added to experimental diet con-

tained the following (g/kg DM): Potassium phosphate (monobasic) 16.68 ; calcium bicarbonate, 5.6; calcium acetate, 20.60; magnesium citrate (dibasic) 7.47; sodium chloride, 5.01 ; cobalt chloride, .0003 ; ferrous sulfate, .2702; cupric sulfate, .0250; magnanese sulfate, .061 ; zinc sulfate, .098 ; potassium iodine, .0002 ; sodium selenite, .0002.

eAmount supplied, (mg/kg DM): vitamin A-acetate, 27.1; vitamin D3, 1.62 ~-tocopherol acetate (D-L), 447.5; niacin 3.5; Ca-pantothenate, 19.9; riboflavin, 8.8; pyridoxine, 8.8; thiamin, 8.8; folic acid, .68; biotin, .14; vitamin B~2 , .11; menadixone sodium bisulfite, .11 ; inositol, 361 ; para-amino benz- oic acid, 18.1; ascorbic acid, 226.0; chlorine chloride, 4700.

percent of the dry matter , were 0, 1.29, 1.82, and 3.04. Exper iment 3 was designed to test the essentiali ty o f meth ion ine and to ascertain the dietary meth ion ine required by the calf. Amoun t s o f me th ion ine were 0, 1.25, 1.82, and 2.45% of the dry matter . During these two experiments , diets were made isoni t rogenous by added or removed glutamic acid f rom the test diet. There were three phases to Exper iments 2 and 3. During the prel iminary phase, the calf was switched f rom the test diet to one o f the diets containing a graded a m o u n t of the specific amino acid. Af te r the calf had received this diet for 3 days at 10% of body weight , metabol ic

measurements were taken for 5 consecut ive days. Measurements were body weight changes, n i t rogen balance, and free amino acid in plasma.

Composition and Formulation of Diets

The exper imenta l diet (Table 1) was pat- te rned after the compos i t ion of whole milk. Milk protein and bu t te r fa t were replaced by a crystal l ine amino acid mix tu re and lard. Leci- thin was added as an emulsif ier to increase ut i l izat ion o f lard by calves, and sodium bicar- bona te was a buffer ing agent to neutral ize acidi ty o f the exper imenta l diet. E t h o x y q u i n was an ant ioxidant . Mixtures of minerals and vi tamins were fo rmula ted f rom data compi led by NRC (9).

Arginine, hist idine, Ieucine, isoleucine, tysine, meth ionine , phenylalanine, threonine, t ryp to- phan, and valine have been indicated essential for growing animals (6, 12). Thus, these 10 amino acids were considered essential for growth in the calf and were included in the test diet. Three nonessent ia l amino acids, proline, tyrosine, and glycine, were added also in the same amounts as in whole milk. L-glutamic acid was added to bring the total n i t rogen con ten t of the exper imenta l diet to that of whole milk.

The exper imenta l diet was prepared by dissolving lactose in water fo l lowed by addi t ion o f both lard and leci thin and raising the tem- perature to approx imate ly 27 C. Af ter the addi t ion o f minerals, the solut ion was di luted to a concen t ra t ion o f 14.6% dry mat ter . The tempera ture then was raised to 38 C for 15 min, and after cooling, the vi tamins were added. The mix tu re was pasteurized at 75 C for 5 min and homogen ized at a pressure of 1,136 to 1,363 kg/6.45 cm 2. The pasteurized, homog- enized mix ture was stored in a freezer.

In preparat ion of the diet for feeding, the mix tu re was defrosted, and the quan t i ty required for each meal was removed and supplemented with desired amount s of the amino acid mix tu re and sodium bicarbonate . Again the final solut ion was adjusted to a concen t ra t ion o f 14.6% dry mat te r wi th warm water. The l iquid test diet was mixed well and hand-fed via a nipple bot t le to each calf twice daily.

Sampling and Chemical Analyses

Daily ur ine ou tpu t was col lected f rom each

Journal of Dairy Science Vol. 63, No. 3, 1980

AMINO ACID NUTRITION IN THE CALF 443

calf by suspending a plastic sheet beneath the metabolism cage. Urine was allowed to filter through glass wool into a plastic bott le containing about 15 ml of concentrated HC1. One-hundredth of the daily urine output was removed and composited during the 5-day collection. The urine samples were stored under toluene in a frozen state until analyzed. A plastic bag at tached to strips of cloth glued to the rear of the calf was used to collect feces. The daily fecal output was weighed, mixed, and a sampIe removed. Representative samples of feed, feces, and urine were analyzed for dry matter , total nitrogen, and ether extract as outl ined by AOAC (1).

On the day following conclusion of collec- tions of feces and urine in Experiment 1, jugular blood samples were taken from each calf to study the effects of diet and time of sampling on patterns of free amino acids in plasma. Shortly before the morning feeding a zero-time blood sample was drawn, and the calf was fed one-half of its daily ration; blood samples were taken at 20 and 40 rain and at hourly intervals for 5 h.

The day following termination of Experi- ments 2 and 3, a zero-time blood sample was taken prior to the morning feeding, and the calf then was fed one-twenty fourth of its daily feed allowance at hourly intervals for 4 h. A single blood sample was taken at h 5 following the initial feeding. This technique of feeding and sampling was adopted to obtain a more representative picture of amino acid availability and uti l ization (5).

About 40 ml of blood were taken from the external jugular vein at each bleeding. The blood was collected in a 50-ml plastic centrifuge tube containing heparin. Plasma was obtained by centrifugation at approximately 8,000 x g for 20 min. Twenty milliliters o f plasma were transferred to a 50-ml plastic centrifuge tube which contained 1 g of sulfa- salicyclic acid dissolved in 2 ml of an aqueous solution containing 2.5 pmoles of norleucine. This mixture was shaken thoroughly and allowed to stand in an ice bath from 2 to 4 h before submission to centrifugation at 31,000 x g for 30 min. The supernatant was recen- trifuged at 31,000 x g for 30 min to ensure that all undesired components were removed. Ten milliliters of supernatant were mixed with 2 ml of polyethylene glycol, (carbowax 400) in

a plastic vial and stored under liquid nitrogen until analyzed for free amino acids by a Phoenix amino acid analyzer model K-8000 C.

RESULTS A N D DISCUSSION

Experiment 1 : Evaluation of Experimental Diet

Body Weight Changes and Digestibilities of Diets. Table 2 shows changes in body weight of the calves and digestibilities of dietary components. All animals gained weight during the exper iment ; however, calves fed the control diet gained significantly more weight (P<.01) than those fed the test diet. Although the mean weight gain on the test diet was significantly less than on whole milk, daily gain was considered acceptable and within the range of gains (330 to 650 g/day) reported by NRC (9).

Apparent digestibilities of dry matter, fat, and total nitrogen were less (P<.05) for the test diet than for the control diet. The reason for the low apparent digestibility of nitrogen when supplied as free amino acids is not known. However, a number of factors may have been responsible for the results. First, a rapid passage of the amino acids to the absorption site may have saturated the system, allowing the escape of amino acids on down the gastrointestinal tract. Second, there may have been compet i t ion at the absorption sites between specific amino acids and/or groups of amino acids which may have led to lower absorption rates for some amino acids (11). Third, since amino acid transport across the gut wall appears to be by active transport , a shortage of energy at the absorption site at the time of greatest amino acid availability may have lead to a decrease in absorption. Support for these explanations, either from the present study or other studies, is unlimited.

The apparent digestibility of the lard (ether extract) of the test diet was substantially less than that of the fat in whole milk. It generally is recognized that the digestibility of lard by young calves is less than that of but terfat (10). The protein content of the diet also influences digestibility o f dietary fat (2).

Nitrogen Balance. Data on nitrogen balance of the calves consuming control and test diets are in Table 3. Mean fecal and urinary nitrogen losses for the calves on the test diet were greater (P<.05) than the losses when they were


444 TZENG AND DAVIS

TABLE 2. Body weight gains on the control and test diets and apparent digestibility coefficients of dietary components.

Apparent digestibilities Calf Body weight Dry Ether no. Diet a gain matter extract Nitrogen

(g/day) (%). SE X SE X SE X SE

1 Control 748 68 b 96.6 1.6 98.7 .3 93.9 2.3 Test 499 70 87.5 2.3 75.8 1.8 81.5 4.8

2 Control 737 34 93.1 1.7 94.4 1.8 90.0 1.0 Test 427 27 81.8 1.7 82.0 2.8 86.4 1.6

3 Control 617 18 95.4 .8 98.4 .60 94.3 1.9 Test 476 23 87.3 .8 80.7 4.4 86.4 1.4

Mean Control 700 33 c 95.0 .9 c 97.2 1.0 c 92.7 1.2 d Test 444 23 85.5 1.4 79.5 1.9 84.8 1.7

acontrol diet-whole milk; test diet-semipurified diet in which crystalline amino acids supplied the total nitrogen.

bEach entry in table represents mean of two observations ± SE. Cpaired-test showed significance between diets at P<.01. dDifferent at P<.05.

fed the cont ro l diet. Nitrogen re tent ion indices (g/day or % of absorbed N) were lower (P<.05) for calves on the test diet than those on the control diet. Al though the mean ni t rogen re tent ion by the calves on the test diet was only two-thirds of that o f calves on the milk diet, it compares favorably with ni t rogen re tent ion for calves fed diets conta ining e i ther dried skim milk or casein as the sources o f ni t rogen (3, 4).

Plasma Free Amino Acid (PFAA). Concen- t rat ions in plasma of 10 free amino acids at seven t imes after feeding are summarized in Tables 4 and 5. In calves consuming the cont ro l diet, all free amino acids in plasma increased in concen t ra t ion during the first post feeding plasma sample (Table 4). However , increases were relatively small compared to increases when the test diet was fed (Table 5). Since the plasma concent ra t ion of a part icular amino acid represents a balance be tween absorpt ion and uti l izat ion, one might conc lude that a more adequate balance existed be tween availability and ut i l izat ion of amino acids f rom the control diet than for the test diet. Also, since no single amino acid showed a marked increase in plasma concent ra t ion , availability o f the amino acids f rom the milk proteins was in reasonably close balance with the needs o f the animal for

pro te in synthesis. In contrast to the small increases in P F A A of

calves receiving the cont ro l diet, the P F A A of calves on the test diet exhibi ted large increases immedia te ly af ter feeding. In mos t cases, the increases for all amino acids were several fold above the base line (zero- t ime sample). The greatest increase was with the amino acid lysine which increased in concent ra t ion by a factor of 4.8 t imes above zero time. Methionine and phenylalanine showed the smallest increases in plasma concent ra t ion . The rapid rise in amino acids of plasma in calves on the test diet, compared to the animals on the cont ro l diet, indicates tha t absorpt ion of the amino acid f rom the gut occurred rapidly. The rapid increase in plasma of amino acids immedia te ly after feeding possibly resulted in inefficiencies in ut i l izat ion o f amino acids for prote in synthesis as exempl i f ied by higher losses o f ur inary nitrogen. The ineff iciencies could have resulted f rom lack o f available energy for incorpora t ion o f the amino acids into prote in at peak absorp- t ion.

The results o f this s tudy poin t to a serious p rob lem in a t t emp t s to for t i fy a diet conta ining pre formed prote ins with a specific amino acid to improve ni t rogen ut i l izat ion. Free amino



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Experiment 2: Lysine Requirement of the Young Calf

Growtb. Results of feeding graded amounts of lysine-HC1 (0, 1.29, 1.82, and 3.04% of dry matter of the diet) on changes of body weight in calves are in Table 6. When the diet was devoid of lysine (0), the calves lost on the average 302 g of body weight per day. These data, plus those for nitrogen balance, demonstrate essentiality of lysine for the young calf, a finding which was not unexpected but which had not been demonstrated. Positive response in body weight gain was observed when the diet contained 1.29% lysine; however, the maximum response was at 1.82% of dietary dry matter . A higher amount (3.04%) did not cause a further increase in weight gain.

Nitrogen Balance. Data from feeding graded amounts of lysine on nitrogen balance are in Table 6. Calves fed the diet devoid of lysine were in negative nitrogen balance to the extent of 3.63 g/calf/day. Including lysine in the diet at 1.29% of dry mat ter resulted in a significant positive retention of nitrogen (mean 7.14 g/calf/day) and increasing lysine to 1.82% further increased nitrogen retention (8.80 g/calf/day). However, increasing lysine to 3.04% of the dietary dry mat ter did not increase nitrogen retention above that obtained when lysine was at 1.82% of the dry matter. Thus, these data substantiate those for gain in body weight. Both measurements indicate that 1.82% lysine in the diet is sufficient to optimize weight gain and nitrogen balance in the young calf.

Plasma Free Amino Acids. Lysine in plasma when calves were fed diets containing various percents of lysine are in Table 6. Lysine con- centrat ion was extremely low (32/amol/l i ter) in plasma of the calves fed the diet devoid of


446 TZENG AND DAVIS

TABLE 4. Free amino acid in jugular plasma of calves on control diet (whole milk) a.

Po stfeeding

Min Amin6 acid 0 20 40 60 120 180 240

(#mol/liter) Threonine 101 101 96 88 88 93 107 Valine 317 330 334 279 279 294 290 Methionine 42 47 47 42 47 37 40 Isoleucine 110 108 101 105 118 100 92 Leucine 194 174 163 180 168 156 141 Tyrosine 44 55 51 72 65 55 47 Phenylalanine 66 81 77 111 93 101 78 Lysine 98 100 85 90 104 98 95 Histidine 51 73 58 58 73 65 61 Arginine 73 95 89 65 68 67 73

aMeans of three calves.

lysine. Adding lysine-HC1 to the d ie t at 1 .29% of dry m a t t e r increased lysine in p lasma (128 /amol / l i te r ) ; however , a f u r t he r increase of lysine in the diet to 1.82% of d ry m a t t e r resul ted in on ly a small increase in the p lasma c o n c e n t r a t i o n o f this amino acid (134 /amol/ l i ter). Increasing lysine in t he diet above 1.82% o f the dry m a t t e r led to a m a r k e d increase in p lasma c o n c e n t r a t i o n s o f iysine (501 /amol/ l i ter). Thus, avai labi l i ty and u t i l i za t ion of lysine were in a ba l anced s ta te at or near d ie ta ry

lys ine o f 1 .82% of the d ry ma t t e r . D ie ta ry lys ine above 1.82% o f the dry m a t t e r appea red to be in excess of needs of calves as ev idenced by the h igh c o n c e n t r a t i o n of lysine in plasma.

F r o m da ta o f th i s s t udy ( b o d y weigh t gain, n i t rogen balance, and lysine in p lasma) , an a t t e m p t was m a d e to es tabl ish the d ie t a ry r e q u i r e m e n t for lysine. Because of the l imi ted n u m b e r o f animals , conc lus ions can be on ly ten ta t ive . Re la t ionsh ips o f rate of b o d y we igh t gain, n i t rogen r e t en t ion , and p lasma lysine to

TABLE 5. Free amino acid in jugular plasma of calves on experimental diet a.

Postfeeding

Amino acid 0 20 40 Min 60 120 180 240

(#tool/liter) Threonine 98 295 251 196 196 189 161 Valine 185 631 530 448 417 530 464 Methionine 37 80 77 84 78 50 51 lsoleucine 103 284 219 180 133 106 101 Leucine 175 498 391 308 247 196 174 Tyrosine 54 144 115 109 95 53 55 Phenylalanine 85 163 153 141 112 102 83 Lysine 86 412 350 243 256 173 122 Histidine 39 140 115 84 72 75 72 Arginine 78 256 217 174 130 91 97

aMean of three calves.



TABLE 6. Effect of dietary lysine on body weight gain, nitrogen retention, and lysine in plasma in the young calf.

Mean Dry Body Number of Lysine-HC1 Lysine body matter weight Lysine animals in diet intake weight intake gain N-retention in plasma

(% of DM) (g/day/ (kg) (g/day/ (g/day) (g/day) (#mol/ kg BW) (kg BW) liter)

3 0 0 26.7 14.6 -302 -3.63 32 2 1.29 .21 44.1 16.3 454 7.14 128 2 1.82 .28 46.6 15.6 680 8.80 134 2 3.04 .47 46.0 15.6 680 8.76 501

lysine intake per day per kg of body weight are in Figure 1. By regression analysis, lines were fitted to the data points to establish the intake of lysine-HC1 necessary for opt imum response. An intake of .27 g/day/kg body weight gave opt imum responses in body weight gain and nitrogen retention. However, for plasma lysine

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Figure 1. Relationship between dietary lysine and gain in body weight, nitrogen retention, and lysine concentration in plasma.

as an index of requirement (17, 18) a slightly higher .31 g/day/kg body weight was observed. Lysine requirement of the young calf is within the range .27 to .31 g/day/kg body weight.

Experiment 3: Methionine Requirement of the Young Calf

Growth. Body weight changes of calves in which graded amounts of D-L methionine (0, 1.25, 1.82, and 2.45% of dry matter) were fed are in Table 7. At zero intake, the calves lost an average of 189 g body weight/day. These data demonstrate the essentiality of methionine for the young calf. Above 0% of methionine in the diet, body weight gains were observed. The maximum average daily gain was for methionine 1.82% of the dietary dry matter. Thus, the quantitative requirement for methionine is met at 1.82% of the dietary dry matter of the diet.

Nitrogen Balance. The results of the nitrogen balance trials for calves fed diets containing four graded percents of methionine also are in Table 7. A mean negative nitrogen balance of 4.59 g/day was obtained on the three experimental calves fed the diet devoid of methionine. These data, combined with those of body weight gain, confirm the essentiality of methionine for the young calf. Including methionine in the diet at 1.25% of the dietary dry mat ter resulted in a significant positive retent ion of nitrogen (10.26 g/day/calf). A further increase in nitrogen retention (12.1 g/day/calf) was obtained when methionine was increased to 1.82% of the dietary dry matter. In agreement with results for weight gain, nitrogen retent ion was opt imized at dietary methionine of approximately 1.80%.

Plasma Free Amino Acid, Free methionine


4 4 8 TZENG AND DAVIS

TABLE 7. Effect of dietary methionine on body weight gain, nitrogen retention, and methionine in plasma in the young calf.

Mean Dry Body Number of D-L-methionine Methionine body matter weight Methionine animals in diet intake weight intake gain N-retention in plasma

(% of DM) (g/day/ (kg) (g/day/ (g/day) (g/day) (#mol/ kg BW) kg BW) liters)

3 0 0 25.8 14.7 -189 -4 .59 28 2 1.25 .18 49.3 14.6 596 10.26 109 2 1.82 .27 50.5 15.0 792 12.10 630 2 2.45 .36 50.7 14.8 769 12.26 888

in plasma is shown in Table 7. Plasma meth- i on ine concentra t ion in the calves fed methionine was extremely low. This is about the same as that in calves (Experiment 1; Table 5) subjected to a fast overnight (37 /~mol/liter). The addition of methionine to the diet at 1.25% of dry matter increased plasma methionine. However, when methionine in the diet was increased to 1.82% of the dry matter, the increase in the plasma meahionine was marked (630 /amol/liter). Methionine requirement, as a percent o f the dietary dry matter , l ies b e t w e e n 1.25% and 1.82%. The e x t r e m e l y high concen- trat ion of plasma methionine (888 / lmol / l i t er ) w h e n the diet c o n t a i n e d 2.45% m e t h i o n i n e , ref lects an over supply o f m e t h i o n i n e in rela- t ion to needs o f the calf.

As for lysine, an a t t empt was made to use the data on weight gain, nitrogen balance, and methionine in plasma to establish the dietary requirement for methionine. The relationship between rate of body weight (g/day/kg initial body weight) and methionine in the diet are in Figure 2. Dietary m e t h i o n i n e o f 1.65% was optimum for gain in body weight of the c a l l This is the requirement of D-L methionine in the absence o f cys t ine . The relat ionship be- t w e e n m e t h i o n i n e and cys t ine needs to be researched in the young calf as well as the e x t e n t to whc ih the cal f can ut i l ize D-methionine.

The relationship between nitrogen retention (g/day/kg body weight) and methionine in the diet (% o f dry matter) is in Figure 2. The breakpoint was at dietary methionine of 1.65%. This agrees with the point for body weight gain as an index of methionine requirement of the calf.

Figure 2 illustrates the relationship between free methionine in plasma and methionine in the diet. Plasma methionine concentration in calves fed the test diet containing .6% meth-

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ionine was included in the calculations. Under the conditions of this experiment, the mini- mum requirement for methionine by the young calf by methionine in plasma as the criterion was 1.15% of dry matter of the diet.

DISCUSSION

Both the dietary lysine and methione requirements estimated from body weight gain and nitrogen balance agreed while the requirement estimated by the PFAA technique was lower. More observations between 1.82 and 2.5% of dietary lysine and between .6 and 1.82% dietary methionine are needed to define more precisely the requirements for these amino acids by the PFAA technique.

During the course of this study Williams and Smith (16) reported on the methionine requirement of the preruminant calf. Their approach was to supplement a synthetic milk diet with varying amounts of L-methionine and to measure the effect upon methionine and urea concentrations in plasma. Using these techniques they found the methionine requirement ranged between 3.9 and 4.5 g/day for calves weighing 50 to 60 kg and growing at .25 kg/day. These workers considered that the cystine intake (.3 g/day) by the calves was too low, thereby increasing the demand for methionine. The total sulfur amino acid requirement was 4.2 to 4.9 g/day or .23 to .26 g/day/kg W "73. Expression of our data for methionine, or possibly more accurately as total sulfur amino acids, on the basis of metabolic body weight (kg W "73) indicates a daily requirement of approximately .65 g/kg W "vs. This is considerably higher than found by Williams and Smith (16) and Williams and Hewitt (15) and possibly results from 1) the use of D-L methionine in our study, 2) differences in the growth rate of the calves (.25 kg/day v s . . 8 kg/day), and 3) differences in apparent digestibility of nitrogen of the diets fed. Adjusting our estimates of the total sulfur amino acid requirement for the above differences between the two studies results in close agreement (.21 to .27 g/day/kg W -73 vs . .23 to .26 g/day/kg W -73). The assumptions in such calculations are 1) D-methionine isJnot used by the calf to meet its requirement for sulfur amino acids. This reduces the available methionine by 50%. 2) Digestibility of methionine was 85%, obtained for the apparent digestibility of nitrogen when

the test diet was fed (Experiment 1). The apparent digestibility of nitrogen for our test diet (85%) was considerably lower than that found by Williams and Hewitt (15) (97%) when calves were fed a diet similar to that used by Williams and Smith (16) in which methionine requirements were established. Thus our values for total sulfur amino acids were lowered by an additional 12%.

Foldager et al. (7) estimated the methionine requirement of the young calf to be in the range of 2.75 to 2.95 g/16 g N or .23 to .26 g/day/kg W "73. It is difficult to understand these estimates since methionine in the diet was not significantly related to body weight gain or to nitrogen balance of the calves. Nor did methionine concentration in the plasma follow the typical pattern showing a distinct breakpoint beyond which increases in methionine in the diet resulted in a linear increase in the concentration of methionine in plasma.

The estimated lysine requirement in this study, 12.4 to 14.3 g/day (.70 to .81 g/kg W -7s) agrees with those reported by Foldager et al. (7) (12.6 g/day or .78 g/kg W -7s ). Our estimates are higher than those reported by Williams and Hewitt (15). Adjusting ours for differences in rate of gain (.7 kg/day vs . . 25 kg/day) and apparent digestibility of nitrogen (85% vs. 97%) would reduce considerably the discrepancy in estimates for the requirement of iysine.

REFERENCES

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5 Bravo, F. O., R. J. Meade, W. L. Stockland, and J. W. Nordstrom. 1970. Reevaluation of the isoleucine requirement of the growing pig: Plasma free isoleucine as a response criterion. J. Anita. Sci. 31:1137.

6 Dudley, W. A., D. E. Becker, A. L. Jensen, S. W. Terrill, and H. W. Norton. 1962. Crystalline amino acid mixture as the sole source of nitrogen for the baby pig. J. Anita. Sci. 31:639.

7 Foldager, J., J. T. Huber, and W. G. Bergen. 1977.


4 5 0 TZENG AND DAVIS

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9 National Research Council. 1971. Nutr ient requ i rements o f domets ic animals. 3. Nutr ient requ i rements o f dairy cattle. 4th rev. ed. Nat. Acad. Sci., Washington, DC.

10 Radastits, O. M., and J. M. Bell. 1970. Nutr i t ion o f the pre ruminant dairy calf with special reference to the digestion and absorpt ion o f nutr ients . A review. Can. J. Anim. Sci. 50:405.

11 Reiser, S., and D. A. Christiansen. 1969. A cross inhibi t ion o f basic amino acid t ransport by neutral amino acids. Biochem. Biophys. Acta. 83:611.

12 Rose, W. C. 1938. The nutri t ive significance of the amino acids. Physiol. Rev. 18:109.

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University Park. 14 Roy, J. H. B. 1974. Problems in the nutr i t ion o f

the p re ruminan t calf. Proc. Nutr. Soc. 33:79. 15 Williams, A. P., and D. Hewitt . 1979. The amino

acid requirements o f the pre ruminant calf. Br. J. Nutr. 41:311.

16 Williams, A. P., and R. H. Smith. 1974. Concen- t ra t ions o f amino acids and urea in the plasma o f the p re ruminan t calf and es t imat ion o f the amino acid requirements . Br. J. Nutr. 33:149.

17 Young, V. R., K. Tontisirin, I. Ozalp, F. Laksh- manan , and N. S. Scrimshaw. 1972. Plasma amino acid response curve and amino acid require- men t in young men: valine and lysine. J. Nutr . 102:1159.

18 Z immerman , R. A., and H. M. Scott. 1965. Inter- relationship of plasma amino acid levels and weight gain in the chick as influenced by subopt imal and superopt imal dietary concent ra t ions o f single amino acids. J. Nutr. 87:13.


Amino Acid Nutrition of the Young Calf. Estimation of Methionine and Lysine Requirements

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