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R. H. COOK AND F. H. BIRD
O'Connor, T. M., 1966. Cell dynamics in the intestine of the mouse from late fetal life to maturity. Am. J. Anat. 118: 525-536. Reyniers, J. A., M. Wagner, T. D. Luckey and H. A. Gordon, 1960. The White Wyandotte bantam and White Leghorn chicken. A survey of germ-free animals. Lobund Report #3, Notre Dame University Press, Notre Dame, Indiana. Sprinz, H., 1962. Morphological response of intestinal mucosal cells to enteric bacteria and Asiatic cholera.
Fed. Proc. 21: 57-64. Sprinz, H., G. J. Kendal, R. E. Dammin, H. Horowitz, H. Schneider and S. B. Formal, 1961. The response of the germ-free pig to oral challenge with E. coli and Shigella flexneri. Am. J. Path. 39: 681-695. Thrasher, J. D., and R. C. Greulich, 1965. The duodenal progenitor population. II. Age related changes in size and distribution. J. Expt. Zool. 159: 385-396.
PAUL V. TWINING, J R . , O W E N P . THOMAS, ERNEST H . BOSSARD AND JAMES L . NICHOLSON
Department of Poultry Science, University of Maryland, College Park, Maryland 20740 (Received for publication February 27, 1973)
ABSTRACT Six experiments were conducted with broiler cross male chicks to determine the available lysine requirement during the 7-9 week growing period. The birds were reared from day-old to 7 weeks of age on commercial type broiler rations. At 7 weeks of age the birds were group-weighed by pens and the pens ranked according to average weight and blocked on this basis. The treatments were randomly assigned within each block. Graded levels of lysine from L-lysine • HC1 were added to lysine deficient basal rations which had been formulated based on available lysine values of 0.26, 3.09, 0.71 and 1.04% for corn, soybean meal, corn gluten meal and sesame meal, respectively. Using regression analysis the minimum available lysine requirement was calculated for this period. The minimum lysine requirement for male broilers during this period was 0.68% under optimum growing conditions. In one experiment the requirement appeared to increase under summer stress conditions. When expressed as a function of energy this would be 0.201% available lysine per mega Calorie M.E. per kg. of diet. POULTRY SCIENCE 52: 2280-2286, 1973
INTRODUCTION IRD (1953) reported that the lysine requirement of 8 week-old Rhode Island Red chickens for optimum growth and feed efficiency was 0.72% of the total ration. He used a 16% protein diet which contained 33.87% sesame meal as a low lysine basal ration. The energy content of the diet was not given. Schwartz et al. (1958) used a diet containing 32% sesame meal in their lysine studies. They suggested that with a diet containing 975 Calories of productive energy per pound (2145 Calories P.E. /kg.), the lysine
B
1. Scientific Article No. A1817. Contribution No. 4617 of the Maryland Agricultural Experiment Station. Department of Poultry Science.
requirement for birds 6-8 weeks of age would be 0.9% of the diet. For birds 8-11 weeks of age a suggested requirement of 0.8% of the diet was given. Their studies showed that changing the energy concentration of the diet altered the lysine requirement at a rate of 0.07% per 100 Calories of productive energy. Combs and Nicholson (1964) proposed amino acid-energy ratios for broiler rations by growing periods. For the period of 5-8 weeks of age lysine levels of 0.71 and 0.78% per mega Calorie metabolizable energy per pound (0.32 and 0.35% per mega Calorie M.E./kg.) were recommended for the winter and summer, respectively. After 8 weeks of age 0.60 and 0.67% lysine per 1000 kilocalories metabolizable energy per pound (0.27
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The Available Lysine Requirement of 7-9 Week Old Male Broiler Chicks1
BROILER AVAILABLE LYSINE REQUIREMENT
Combs (1970) related the lysine requirement to the ambient temperature. Expressed as the available lysine requirement per mega Calorie metabolizable energy per kg. for broilers 6-7 weeks of age, the values are 0.33, 0.34 and 0.35% for ambient temperatures of 70, 80 and 90° F . , respectively. The requirement for broilers over 8 weeks of age is 0.28, 0.29, 0.30 and 0.31% for ambient temperatures of 60, 70, 80 and 90° F., respectively. Bornstein (1970) reported the lysine requirement for broilers during the 5-8 week period as 0.92% of the diet, or 5.0% of the dietary protein, or about 33.0 gm. per mega Calorie M.E. The corresponding values for the 8-10 week period were given as 0.76 and 4.4% and 2.5 gm., respectively. EXPERIMENTAL PROCEDURE Experiments were designed to measure the lysine requirement of the broiler chicken during the period from 7-9 weeks of age. In all the studies broiler type birds were reared from day-old to 7 weeks of age on commerical type broiler rations. Broiler starter crumbles were fed for the first 4 weeks and broiler finisher pellets were fed for the period of
4-7 weeks of age. At 7 weeks of age all birds were group weighed by pens. The average weight of the birds for each pen was calculated and the pens were then ranked according to the average weight and blocked on this basis. The treatments were randomly assigned within each block. All dead birds were weighed during the experimental period and adjustments made for mortality when calculating the feed conversion. The body weight gain and feed conversion data were statistically analyzed by analysis of variance and by the Student-Newman-Keuls' multiple range test (Sokal and Rohlf, 1969). In manufacturing the feed, a basal mix was prepared which contained all of the ingredients except glutamic acid and L-lysine HC1. Various levels of L-lysine HC1 were added to the basal. Sufficient glutamic acid was added in combination with L-lysine HC1 so that the nitrogen content of the rations for all treatments remained constant. After adequate mixing, the ration for each treatment was pelleted in a commercial pellet mill (Spout-Walden) at a temperature of 88° C. The calculated available lysine levels of the basal rations for the first three studies were 0.60, 0.553 and 0.554 (Table 1., Exps. 1, 2 & 3). The fourth trial was conducted at the University of Maryland sub-station at Salisbury, Maryland. This facility is divided in the center by a feed room and consists of 32 test pens containing 150 birds each. Forty-eight hundred Hubbard x Arbor Acre Line-60 male crosses were assigned to the 32 test pens and raised to 7 weeks of age. At this point four pens were discarded from the study. In order to obtain a greater growth response to graded levels of lysine a basal ration (Table 1, Exp. #4) was formulated to contain 0.47% available lysine based on available lysine values of 0.26, 3.09 and 0.71% for corn, soybean meal and corn gluten meal, respectively. The basal ration supplemented with L-tryptophane was calculated to contain at
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and 0.35% lysine per 100 kilocalories M.E./kg.) were recommended levels for winter and summer, respectively. The values were based on diets formulated using the then current Maryland feed composition tables. Combs (1968) updated his proposed lysine requirements based on revised nutrient composition data of feed ingredients presented at that time. Two trials were conducted to evaluate the lysine requirement of broilers from 4-8 weeks of age. Using regression equation procedures a value of 0.74% available lysine per mega Calorie of metabolizable energy per pound (0.33% available lysine per mega Calorie M.E./kg.) was calculated for this period. For the period after 8 weeks an extrapolated value of 0.64% available lysine per therm M.E. per lb. (0.29% available lysine per therm M.E./kg.) was postulated.
2281
2282
TWINING, JR. , THOMAS, BOSSARD AND NICHOLSON
TABLE 1.—Experimental rations
Ingredients
%
%
%
%
%
%
%
%
62.73
64.35
66.24
65.76
69.40 13.08 11.42
64.71 18.72 4.72 4.08
62.43
—
66.41 18.70 7.03 0.86
1.93 3.61
1.88 4.57
1.87 5.36
—
—
—
22.50 8.50
22.75 7.25
22.25 7.25
26.50 3.59
1.87 2.00 1.50
— 2.00
1.96 1.25
1.97 1.00
—
— 2.51 23.27 0.76 1.00 5.98 3.00
— 0.43 0.19
1.92 0.85 0.52
0.46 0.27
0.62 0.31
0.14 0.05 0.05 0.03 0.01
0.19 0.05 0.05 0.023 0.01
0.19 0.05 0.05 0.021 0.01
0.20 0.05 0.05 0.029 0.01
—
—
—
0.27
0.28
0.28
0.18 0.05 0.05
0.16 0.05 0.05
0.15 0.05 0.05
0.35 0.36 0.11 0.12 0.05 0.05
—
—
—
—
0.01
0.01
0.01
0.01
3368.00 3369.00 3359.00 3388.00 3388.00 3388.00 3388.00 3366.00 23.86 24.43 21.51 21.51 20.95 24.48 23.71 21.00 0.553 0.554 0.467 0.398 0.444 0.523 0.60 1.03 0.64 0.59 0.60 0.47 0.58 0.60 0.58 0.59 0.98 0.89 0.91 0.95 0.78 0.89 1.00 0.95 0.20 0.22 0.28 0.27 0.22 0.19 0.20 0.25
'To supply the following per kilogram of diet: Vitamin A, 6614 I.U.; vitamin D3, 2205 I.U.; vitamin E, 3.0 I.U.; menadione, 1.46 mg.; riboflavin, 6.6 mg.; pantothenic acid, 11.0 mg.; niacin, 26.5 mg.; folacin, 0.22 mg.; vitamin B12, 13.2 meg.; manganese, 75 mg.; zinc, 40 mg.; copper, 3.0 mg.; iodine, 1.2 mg. In addition ethoxyquin3 was supplied at 0.0220%. 2 Zoalene, Dow Chemical Company, Midland, Michigan. 3 Santoquin, Monsanto Company, St. Louis, Missouri.
least 105% of the currently accepted requirements in all the essential amino acids except lysine (Combs, 1970). This diet had a calculated energy content of 3388 kcal. M.E./kg. L-lysine HCl was added to the basal in increments of 0.074% lysine to give 7 levels of lysine. Average weight gain and feed consumption data were collected after the birds were on test for 10 days. In study 5, Ledbrest x Arbor Acre Line-70 male crosses were reared as in the previous studies. At 7 weeks of age all sexing errors were removed, the number of birds per pen reduced to 45, and the birds group weighed
by pens. A basal ration similar to the one used in trial 4 was employed in this study. L-lysine HCl was added to the basal in increments of 0.116% lysine to give 5 levels of lysine. Three dietary treatments containing TABLE 2.—Available lysine content of basal rations as per chick bioassay Experiment No.
Calculated avail, lysine
1 2 3 5
0.60 0.553 0.554 0.469
(%)
Assayed avail, lysine
(%)
0.588 0.515 0.528 0.455
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Ground yellow corn Sesame meal Corn gluten meal-60% protein Soybean meal-50% protein Fish meal, herring Defluorinated phosphate Stabilized animal fat Whey Meat and bone meal-48% Dicalcium phosphate Limestone Salt Methionine hydroxy analog Choline chloride-50 Vitamin-trace mineral mix1 Coccidiostat 2 L-Tryptophane Ferrous sulfate heptahydrate Calculated Analysis Metabolizable energy, kcal./kg. Protein, % Lysine, % Methionine, % Methoinine plus cystine, % Tryptophane, %
•Exp. s 5 Exp. # l E x p . #2Exp. #3Exp. # 4 Diet A D i e t B Diet C Exp. # 6
BROILER AVAILABLE LYSINE REQUIREMENT
2283
TABLE 3.—Average gains and feed conversions of chicks fed diets deficient and adequate in lysine (College Park). (Experiment I). Gain Feed conversion Lysine 7-8 1/2wks. 7-9 wks. 7-8 1/2wks. 7-9 wks. (% Diet) (kgO (kg?) 0.60 0.530s" 0.776" 2.37a 2.50* ab 0.69 0.562 0.793" 2.21b 2.42ab 0.79 0.565"b 0.803" 2.19b 2.37b 0.88 0.560"b 0.803" 2.17b 2.34b 0.98 0.575ab 0.826" 2.16b 2.32" 1.07 0.586b 0.834" 2.15b 2.33b 'For any column, values with the same superscript to the right are not significantly different at (P < 0.05).
Experiment 6 was conducted to confirm the results of studies 4 and 5 and to test the lysine requirement for males by including a practical type diet (Table 1, Exp. #6) as a positive control. Graded levels of L-lysine HC1 were added to a low lysine basal ration which contained sesame meal (Table 1, Exp. # 5 , Diet A). Vantress x Arbor Acre Line-70 male crosses were reared to 7 weeks of age, at which time all sexing errors were removed, the number of birds per pen reduced to 50, and the birds group weighed by pens. Average weight gain and feed consumption data were
collected after the birds were on test for 10 days and again after 2 weeks. RESULTS AND DISCUSSION During the 7-8-1/2 week period the weight gain for the lowest level of lysine in experiments 1 and 2 (Tables 3 and 4) was significantly different (P < 0.05) from the highest level of added lysine. There were insufficient points to construct a meaningful response curve. In experiment 3 the same trend was apparent (Table 5). The data from experiment 4 are summarized in Table 6. The birds in experiment 4 were on the test diets during a severe 2
TABLE 4.—Average gains and feed conversions of chicks fed diets deficient and adequate in lysine (Salisbury). (Experiment 2). Lysine (% Diet) 0.55 0.62 0.69 0.76 0.83 0.89 0.96 1.03
Gain Feed convs 7-8 1/2 wks. 7-8 1/2 \ (kg.) 0.349" ' 0.396ab 0.364ab 0.389"" 0.403 ab 0.390"" 0.387"" 0.435"
3.64" 3.01" 3.35" 3.09" 3.10" 3.19" 2.95" 2.82"
1 For any column, values with the same superscript to the right are not significantly different at (P < 0.05).
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sesame meal were incorporated in the design for comparison of sesame meal with corn gluten meal as a low lysine ingredient. These rations, diets A, B and C (Table 1, Exp. #5) were formulated to contain 0.40, 0.44 and 0.52% available lysine, respectively, based on an available lysine value of 1.04% for sesame meal which was determined by a chick bioassay as described by Combs et al. (1968). The basal rations employed in experiments 1-3 and 5 were also assayed for available lysine by this method (Table 2). The available lysine values used for the other ingredients were the same as those used in experiment 1. The sesame meal diets had a calculated energy content of 3388 kcal. M.E./kg. Average weight gain and feed consumption data were collected after the birds were on test for 10 days and again after 2 weeks.
2284
TWINING, JR. , THOMAS, BOSSARD AND NICHOLSON
TABLE 5.—Average gains and feed conversions of chicks fed diets deficient and adequate in lysine (College Park). (Experiment 3) Feed conversion 7-8 1/2 wks. 7-9 wks.
Gain Lysine
7-8 1 / 2 w k s .
7-9 wks.
(% Diet) 0.55 0.62 0.69 0.76 0.83 0.89 0.96 1.03
(kg.) 0.499 s ' 0.549" 0.549" 0.535" 0.562" 0.562" 0.558" 0.553"
(kg.) 0.617" 0.676" 0.680" 0.689" 0.712" 0.712" 0.712" 0.703"
2.64" 2.58" 2.50" 2.48" 2.42" 2.43" 2.41" 2.50"
2.79" 2.68" 2.63" 2.53" 2.51" 2.52" 2.51" 2.54"
'For any column, values with the same superscript to the right are not significantly different at (P < 0.05).
TABLE 6.—Average gains and feed conversions of chicks fed diets deficient and adequate in lysine (Experiment 4). Lysine (% Diet) 0.47 0.54 0.62 0.69 0.76 0.84 0.91
Feed conversion Gain 7-8 1/2 wks. 7-8 1/2 wks. (kg.) 0.234"' 0.210" 0.290" 0.317" 0.337" 0.338" 0.345"
4.27"" 4.65" 3.69"= 3.24c 3.12c 3.09° 3.10c
' For any column, values with the same superscript to the right are not significantly different at (P < 0.05).
conversion than those in the west end. Although there is insufficient data to draw any definite conclusion, the results suggest that the heat stress raised the requirement. Regression equations were calculated for the gain and feed conversion during the 7-8-1/2 week period using the data from the treatments on the 5 lowest levels of lysine and are as follows: Period 7-8-1/2 weeks 7-8-1/2 weeks
Criteria gain in body weight (kg.) feed conversion
Regression Equation2 y = 0.406 + 0.593 logX y = 2.273 - 7.018 logX The data from experiment 5 are summarized in Table 7. Regression equations were calculated for the gain and feed conversion during the 7-8-1/2 and 7-9 week periods
6 lowest levels of lysine and are as follows: Period 7-8-1/2 weeks 7-9 weeks 7-8-1/2 weeks 7-9 weeks
Criteria gain in body weight (kg.) gain in body weight (kg.) feed conversion feed conversion
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day heat wave with temperatures reaching into the upper 90's during the day. Some of the birds in the east end of the research house—a windowless fan ventilated unit— died from the heat during this period. No birds were lost from heat in the west end which was completely open and naturally ventilated. Maximum weight gain was obtained in the west end of the house with 0.69% lysine. In the east end, where the birds were stressed more from the heat, 0.76% lysine was required for maximum weight gain. At all treatment levels the birds in the east end gained significantly less (P < 0.01) than those in the west end. The birds in the east end also had a significantly higher (P < 0.01) feed
BROILER AVAILABLE LYSINE REQUIREMENT
2285
TABLE 7.—Average gains and feed conversions of chicks fed diets deficient and adequate in lysine (Experiment 5). Gain Feed conversion Lysine 7-8 1/2wks. 7-9 wks. 7-8 1/2wks. 7-9 wks. (% Diet) (kgO (kgO A 0.40 0.304al 0.372" 3.74a 3.99s b b B 0.44 0.331 0.413" 3.29 3.38c 0.47 0.345b 0.427b 3.36b 3.54b c c C 0.52 0.435 0.544 2.70° 2.77d 0.58 0.476d 0.594" 2.64c 2.70" 0.69 0.499° 0.621de 2.55c 2.64d 0.81 0.513e 0.640* 2.52c 2.60d e de 0^93 0.513 0.626 2^52° 2.61" 'For any column, values with the same superscript to the right are not significantly different at (P < 0.05).
Gain Feed conversion Lysine 7-8 1/2 wks. 7-9 wks. 7-8 1/2 wks. 7-9 wks. (% Diet) (kgO (kg?) 0.40 0.435al 0.576a 2.92° 2.90c 0.49 0.490" 0.612" 2.63" 2.74" 0.58 0.526° 0.671c 2.47"" 2.53a 0.67 0.553d 0.694°" 2.36a 2.46a 0.76 0.572d 0.726e 2.30" 2.37a 0.85 0.581" 0.730e 2.29" 2.38a 1.03 (Table 1) 0.572d 0.717de 2.39s 2.50" (Ex. #6) 'For any column, values with the same superscript to the right are not significantly different at (P < 0.05). Regression Equation* y = 0.669 + 0.922 logX y = 0.839+ 1.173 logX y = 1.528 - 5.167 logX y = 1.479 - 5.752 logX Based on these regression equations the calculated available lysine requirements for maximum growth during the 7-8-1 / 2 and 7-9 week periods are 0.68 and 0.675%, respectively. The requirement for optimum feed efficiency for both of the above periods was 0.65%. When expressed as a function of energy these requirements would be 0.201,
2. Where X is percent lysine in the diet.
0.199, 0.192% lysine per mega Calorie M.E. per kg. of diet, respectively. The lower requirements obtained in experiment 5 as compared to those found in experiment 4 are mainly a reflection of the improved environmental conditions at College Park, where experiment 5 was conducted, versus those at the Salisbury sub-station, where experiment 4 was run. In experiment 5 there were fewer birds per pen, more square feet of floor space per bird, less severe weather conditions and better overall Ventilation. The data from experiment 6 (Table 8) confirmed the findings of studies 4 and 5. Regression equations were calculated for the gain during the 7-8-1 / 2 and 7-9 week periods
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TABLE 8.—Average gain and feed conversion during the withdrawal period with cockerels (Experiment 6).
2286
TWINING, JR. , THOMAS, BOSSARD AND NICHOLSON
using the data from the lowest levels of lysine.
for male broilers during this period under
The equations for feed conversion
optimum growing conditions
were
was 0.68%.
calculated using the 5 lowest levels of lysine
When expressed as a function of energy this
and are as follows:
would be 0.201% available lysine per mega Calorie M.E. per kg. of diet.
Period
Criteria
7-8-1/2 weeks
gain in body
weight
7-9 weeks
(kg.) gain in body
weight
7-8-1/2 weeks
(kg.) feed conversion
7-9 weeks
feed conversion Regression Equation? y = 0.792 + 0.550 logX y = 1.987 - 2.218 logX y = 2 . 1 1 8 - 1.947 logX
Based on these regression equations the calculated available lysine requirements of males for maximum growth and optimum feed efficiency during the 7-8-1/2 and 7-9 week periods are 0.70 and 0.72% respectively. The requirement values for male broilers during the 7-9 week period obtained from these studies are lower than the
figures
reported by Combs (1970) or Bornstein (1970). The minimum available lysine requirement
Bird, F. H., 1953. The lysine requirement of eight-week old chickens. Poultry Sci. 32: 10-13. Bornstein, S., 1970. The lysine requirement of broilers during their finishing period. Br. Poultry Sci. 11: 197-207. Combs, G. F., and J. L. Nicholson, 1964. Testing energy, amino acid and protein level specifications for linear programming of broiler rations. Feedstuffs, 36(34): 17-21,70. Combs, G. F., 1968. Amino acid requirements of broilers and laying hens. Proc. U. of Maryland Nutrition Conf. for Feed Mfgrs., pp. 86-96. Combs, G. F., E. H. Bossard and G. R. Childs, 1968. Improved chick bioassay for available lysine and available methionine. Feedstuffs, 40(8): 36-37. Combs, G. F., 1970. Feed ingredient composition and amino acid standards for broilers. Proc. U. of Maryland Nutrition Conf. for Feed Mfgrs., pp. 81-89. Schwartz, H. G., W. M. Taylor and H. Fisher, 1958. The effect of dietary energy concentration and age on the lysine requirement of growing chicks. J. Nutrition, 65: 25-37. Sokal, R. R„ and F. J. Rohlf, 1969. Biometry. W. H. Freeman & Co., San Francisco, Calif, pp. 239.
NEWS AND NOTES (Continued from page 2275) CONGRESS NOTES The World's Poultry Congress Committee has named the International Congress and Convention Association as the official congress transportation coordinator for travel arrangements outside the continental United States. Official domestic tours are being exclusively arranged and executed by Courtesy Travel Service on behalf of the congress. The representative for the International Congress and Convention Association is: Paul Padro, Vacationland Travel Service, 300 Madison Avenue, New York, New York 10017. Telephone: 212/867-8147.
Questions regarding domestic tours should be directed to: Richard Sykes, Courtesy Travel Service, 1929 K Street, N.W., Suite 400, Washington, D.C. 20006. Telephone: 202/296-8160. The 1974 World's Poultry Congress will be held August 11-16 in New Orleans.
BRITISH COLUMBIA NOTES A. Akinwande, whose research and thesis was supervised by Dr. D. B. Bragg, Department of Poultry Science, University of British Columbia, Vancouver, received a Ph.D. degree at the Spring Convocation.
(Continued on page 2289)
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y = 0.648 + 0.527 logX
REFERENCES