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Sequence in Which Essential Amino Acids Become Limiting for Growth of Chicks Fed Rations Containing Cottonseed Meal1
Sequence in Which Essential Amino Acids Become Limiting for Growth of Chicks Fed Rations Containing Cottonseed Meal1 J. O. ANDERSON AND R. E. WARNICK Animal Science Department, Utah State University, Logan, Utah (Received for publication June 22, 1965)
EXPERIMENTAL
Five different cottonseed meals were used in these experiments. One commercial 1
Approved as Utah Agricultural Experiment Station Journal Paper No. 489.
84
expeller meal and two commercial prepress solvent-extracted meals were included. Prepress solvent-extracted meal B contained a higher protein level and a lower crude fiber level than meal A. Two meals were prepared under similar conditions at the Cottonseed Products Research Laboratory, Texas A & M University, College Station, Texas.2 One was prepared from glanded seed and the other from glandless seed. The crude fiber, moisture, and protein (N X 6.2S) contents, as well as the solubility of the nitrogen in 0.02N NaOH were determined for each lot of meal. These values are given in Table 1. The composition of the basic rations fed is given in Table 2. All rations fed in these experiments contained 19.1 percent protein (N X 6.25). The cottonseed meal level was varied according to its protein content. In all rations based on a given meal, enough of that meal was used to provide 18 percent protein to the ration. All cottonseed meal rations contained an amino acid mixture that provided an additional 1.1 per2
These meals were provided through Mr. A. Cecil Wamble of the Cottonseed Products Research Laboratory, Texas A & M University, College Station, Texas; and Mr. Garlon A. Harper of the National Cottonseed Products Association, Inc., Memphis, Tenn. The latter reported that chemical analysis indicated that the glanded and glandless meals contain the following, respectively: 0.28 and 0.47 percent residual lipids; 3.5 and 4.1 grams of lysine per 16 grams of nitrogen; 0.08 and 0.01 percent free gossypol; and 1.18 and 0.01 percent total gossypol. The meals were produced by direct hexane extraction; a moderate amount of heat was used during processing.
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IMITED amounts of meal prepared ' from glanded cottonseed are used in poultry rations. It has generally been recognized, however, that the protein in that meal is too deficient in lysine for large amounts to be included in commercial broiler rations. Grau (1946) found that lysine is the most limiting amino acid in cottonseed meal protein; additional methionine was also needed to allow the most rapid growth rate. Further supplementation with threonine or tryptophan did not increase growth rate. Jonston and Watts (1964) have reported that lysine and methionine are also limiting in meals prepared from glandless cottonseed. Lysine was found to be more limiting in the glanded than in the glandless meal. This paper reports the results of experiments conducted to determine the sequence in which the essential amino acids become limiting for growth of chicks in semipurified rations based on five different cottonseed meals. The experiments are similar to others recently reported with peanut meal (Anderson and Warnick, 196S). Our calculations indicated that a ration with 18 percent cottonseed meal protein would be deficient in amino acids other than lysine and methionine. These predictions were tested in these experiments.
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AMINO ACID DEFICIENCIES IN COTTONSEED
85
TABLE 1.—Description of cottonseed meals fed
Type of cottonseed meal
Lot no.
Crude fiber Dry matter
("ptfvfiT'fl
Percent of ' air-dry sample 93 40.1
N soluble in 0.02N NaOH Percent 51
1
15.7
1 2
15.2 16.1
92 91
42.0 39.8
71 70
1 2
10.5 8.4
93 92
49.4 52.0
76 73
Prepared at Texas A & M from glanded seed
1
5.4
94
58.0
77
Prepared at Texas A & M from glandless seed
1 2
5.7 6.2
94 93
61.6 59.1
93 91
Expeller Prepress solvent-extracted Source A Prepress solvent-extracted Source B
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TABLE 2.—Composition of the basic rations cent protein. A ration based on soybean meal and methionine was also fed in all Cottonseed Soybean Ingredient experiments except one. The soybean meal meal meal used contained 50 percent protein and 4.0 Gms./kg. percent crude fiber. Cottonseed meal 295-452! — — 380 A sample of the second lot of prepress Soybean meal, 50% protein Corn oil 10 10 solvent-extracted meal B was hydrolyzed Salt 5 5 with 6N HCL for 24 hours. The levels of Dicalcium phosphate, 15 IS _18.5%P certain amino acids in the hydrolysate Limestone flour 10 10 2 0.6 0.6 were determined by the method of Spack- Trace mineral mix 3 1.4 1.4 Antibiotic supplement man et al. (1958). These levels were used Vitamin mix4 80 80 5 — L-lysine HC1 to estimate cottonseed protein content of DL-methionine 1 2 most amino acids as given in Table 3. The DL-isoleucine 1.5 — 2 — tryptophan and cystine estimates were DL-threonine 1 — DL-valine taken from a report by Almquist (1962). L-leucine 1 — 1 — The calculated level of each amino acid Glycine 1,000 1,000 Glucose monohydrate 5 to provided by a ration with 18 percent cot1 The cottonseed meal level varied with the tonseed protein and the level of each essenmeal's protein content; enough meal was used to tial amino acid in the ration found by provide 18 percent protein in the ration. 2 This mix provided the following in gms./kg. Dobson et al. (1964) to have a well balof ration: FeCeHsO,, 0.2; M n S 0 4 - H A 0.25; KI0 3 , anced mixture of the essential amino acids 0.01; ZnCOs, 0.13; and CuS0 4 , 0.01. 3 Provided 31 mgs. of zinc bacitracin/kg. of are also given in Table 3. The latter two ration. 4 sets of values were compared to predict Composition of this mix is given by Dobson which amino acids should be most limiting et al. (1964), This mix was designed to be used in purified rations at a 10 percent level. 6 in a ration with 18 percent cottonseed meal Cerelose, Corn Products Company, New York, N. Y. protein. When this series of experiments was calculations were based on Almquist's started, the amino acid analysis of cotton- (1962) values. When either set of values seed meal had not been done. The original was used, it was predicted that the same
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J. 0. ANDERSON AND R. E. WAENICK
TABLE 3.—Essential amino acid content of cottonseed meal, a ration with 18 percent cottonseed protein, and the balanced ration of Dobson et a]. (1964)
Amino Acid
Gms.116 gms. N 4.0 1.5 3.51 3.4 3.6 6.3 5.0 2.7 1.3 1 5.5 8.3 11.5 4.4 21.0 9.0 3.8 3.9 4.4
Percent of ration 0.72 1.15 0.27 0.38 0.63 0.61 0.65 1.13 0.90 0.49 0.23 0.99
0.73 0.80 0.78 1.30 0.95 0.43 0.20
1.49 2.07 0.79 3.78 1.62 0.68 0.70 0.79
1.33 1.28
1 Values for cystine and tryptophan were taken from Almquist's (1962) table.
amino acids would be limiting in a ration with 18 percent cottonseed meal protein. The amino acid mixture added to the basic cottonseed meal ration fed in most experiments contained the six strictly essential amino acids predicted to be most limiting in a ration with 18 percent cottonseed protein. This mixture is termed the "complete" mixture in this paper. Valine was not included in the amino acid mixture of the basic ration fed in the first two experiments conducted with the expeller cottonseed meal. Glycine was added to all rations based on cottonseed meal. In each experiment with a given cottonseed meal, the basic ration and five or six other rations based on the same lot of cottonseed meal were fed. Each non-basic ration had one of the strictly essential amino acids removed from the complete amino acid mixture. More glycine was added to
RESULTS AND DISCUSSION
The results obtained are presented in Table 4. Lysine was found to be the most limiting amino acid in all of the commercial cottonseed meals and in the meal prepared in the laboratory from glanded seed. This was expected and comparable results
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Lysine Methionine Methionine & cystine Isoleucine Threonine Leucine Valine Histidine Tryptophan Phenylalanine Phenylalanine & Tyrosine Arginine Glycine Glutamic acid Aspartic acid Alanine Proline Serine
Level provided by Cottonseed 18% meal Cottonseeci Balanced content protein ration ration
these rations to keep the nitrogen level constant. A commercial lysine supplement containing 50 percent lysine was used in some experiments instead of the lysine hydrochloride. Results were similar when either source of lysine was used and have been summarized without regard to the lysine source. Chicks of both sexes from broiler crosses were used in all experiments. They were fed a practical-type ration from two to six days before the experimental rations were fed and were alloted to experimental groups on the basis of body weight just before being started on the experimental rations. Five birds composed each group. The experiments lasted from 14 to 21 days. All groups in an experiment started on experimental rations on the same date and ended on the same date. The birds were housed in electrically-heated, thermostatically controlled battery brooders with raised wire screen floors. Feed and water were supplied ad libitum. At the end of each experiment, the mean weight gain of each group of chicks was expressed as a percentage of the mean weight gain obtained with the basic cottonseed meal ration fed in that experiment. A relative gain/feed ratio was calculated in the same manner. All of the relative values obtained with any one ration were averaged to obtain the relative gains or gain/ feed ratios presented in Table 4. The relative group gains or gain/feed ratios were analyzed by the analysis of variance (Snedecor, 1956).
AMINO ACID DEFICIENCIES IN COTTONSEED
TABLE 4.—Results obtained with five cottonseed meals Amino acid removed from basic ration
Relative gain
Relative gain/feed ratio
Total no. groups
Expeller processed meal—3 experiments 8 100 100 None 1 6 27* 58* Lysine 6 78* 89* Methionine 8 85* 94* Isoleucine 8 91* 98 Threonine 3 92 94 Valine 8 99 101 Leucine Soybean meal & 2 5 99 methionine 114* Prepress solvent-extracted meal A—5 experiments 100 14 None 100 63* 11 Lysine 34* 88* 11 Methionine 70* 91* 13 Isoleucine 79* 92* 14 Threonine 84* 99 14 Leucine 95* 100 11 Valine 100 Soybean meal & 2 117* 13 methionine 99 Prepress solvent-extracted meal B—5 experiments 100 10 None 100 71* 10 Lysine 43* 83* 10 Methionine 68* 92* 10 Isoleucine 85* 96* 10 Threonine 90* 101 10 Leucine 97 99 10 Valine 99 Soybean meal & 109* 10 methionine2 103 Meal prepared from: glanded seed at Texas A 6* M— 2 experiments 100 100 None 4 44* 69* Lysine 4 65* 81* Methionine 4 73* 90* Isoleucine 4 79* 87* Threonine 4 93 97 Leucine 4 103 100 Valine 4 Soybean meal & 2 110* 110* methionine 4 Meal prepared from glandless seed at Texas A &° M—• 3 experiments 100 6 None 100 5 Methionine 73* 87* Lysine 73* 88* 5 93 4 Isoleucine 92 99 4 Leucine 93 4 Threonine 95 99 5 Valine 100 97 Soybean meal & 2 6 methionine 96 101 1 The basic ration fed in the first two experiments of this series did not include added valine. 2 This line gives the results obtained when the soybean meal ration (Table 2) was fed in the same experiments that the cottonseed meal was listed. * Significantly different than gain or gain/feed ratio obtained with cottonseed ration containing the complete amino acid mixture; P<0.01.
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have previously been reported by Grau (1946) who also conducted somewhat similar experiments with semi-purified rations. Many others have found lysine to be the most limiting amino acid in practicaltype rations based on cottonseed meal. The gains obtained when lysine was removed varied from 27 to 44 percent and the gain/ feed ratios varied from 58 to 71 percent of that obtained with the complete amino acid mixture. This suggests that the different meals prepared from glanded seed provided different amounts of available lysine to the chicks. Some of these differences may be due to the random variations in performance noted with this type of test. The low growth rate obtained when lysine was removed from the basic cottonseed meal rations raised a question as to whether the lysine level in the basic rations was adequate to allow' maximum growth rate. In one experiment with the second lot of prepress solvent-extracted meal A, an extra gram of lysine hydrochloride was added per kilogram of the basic ration in one treatment. This increase in lysine did not increase either growth rate or gain/ feed ratio. When lysine was omitted from basic rations using glanded cottonseed meals, the resultant amino acid imbalance probably decreased growth rate. In one experiment all of the essential amino acids were removed from the basic ration. Chicks fed this ration gained more than the chicks fed the ration from which only the lysine was removed from the complete amino acid mixture. Methionine was found to be the second limiting amino acid in all four meals prepared from glanded seed. Grau (1946) also found that methionine is limiting in cottonseed meal protein. Almquist (1962) indicates that cottonseed meal protein contains two grams of cystine per 16 grams of nitrogen. If this is true, then methionine and not the total level of sulphur-contain-
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J. O. ANDERSON AND R. E. WAENICK
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ing amino acids is the limiting factor here. soybean meal ration. The ration made Isoleucine was found to be the third lim- from the meal prepared in the laboratory iting amino acid and threonine the fourth from glanded seed had about the same in each of the meals prepared from glanded crude fiber content as the ration based on seed. Leucine appears to be the fifth lim- soybean meal. The soybean meal ration iting in these meals although the reduction produced a significantly greater gain and in gain noted when leucine was removed better feed efficiency than the basic ration was only statistically significant with prep- made with the laboratory prepared meal. ress solvent-extracted meal A. The overall The meal prepared from glandless cotdifference in gain obtained when leucine tonseed is a much better source of lysine was removed from all four basic rations than any of the four meals prepared from was also statistically significant. glanded cottonseed. Methionine and lysine Valine removal from the different basic appeared to be about equally limiting in rations produced a marked reduction in the meal prepared from glandless cottonperformance only in the third experiment seed. Removal of isoleucine, threonine, or of the experiments conducted with the ex- leucine from this basic ration reduced peller processed meal. Valine was not in- growth rate and gain/feed ratio, but these cluded in the basic ration fed in the first differences were not statistically significant two experiments with this meal. The in these experiments. When lysine, methiodifference noted with valine removal in the nine, isoleucine, or threonine was removed third experiment probably is not real. The from the basic rations containing the two growth rate obtained with the basic ration laboratory prepared meals, the glandless without added valine in the second experi- meal produced the highest relative performent was equal to that obtained with the mance. soybean meal ration. The soybean meal raOne of the experiments with the glandtion was not fed in the first experiment. Our less meal and one with the glanded meal supply of this meal was exhausted after were done at the same time with chicks three experiments, and it was not consid- from the same hatch. Chicks fed the basic ered worthwhile to obtain a new lot for ad- ration with glandless cottonseed meal ditional tests. gained 16.8 grams per day with a gain/ The growth rates obtained with the feed ratio of 0.56, while chicks fed the basic rations made with commercial cot- basic ration with the glanded meal gained tonseed meals were comparable to those 14.5 grams per day with a gain/feed ratio obtained with the ration based on soybean of 0.51. Thus, the differences in absolute meal and methionine, but the soybean meal growth rate and gain/feed ratio were even ration produced significantly better feed greater than the relative differences given efficiency. The rations based on commercial in Table 4. The protein of the meal precottonseed meals contained from 1.5 to 5.8 pared from the glandless seed appears to percent more crude fiber than the ration provide more of at least these four amino based on soybean meal. All of these rations acids than the glanded meal protein. Kuikcontained a rather wide energy/protein en (1952) found that lysine availability ratio. The higher fiber level in the cotton- was greatly reduced by heating cottonseed seed rations probably made the growth meal with gossypol dissolved in cottonseed rate obtained with these rations higher oil. Luecine, isoleucine, valine, threonine, than would have been obtained if the crude and methonine availabilities were also refiber level had equaled that found in the duced by this treatment.
AMINO ACID DEFICIENCIES IN COTTONSEED
When this mixture of seven amino acids was added to the glandless cottonseed meal, the resulting growth rate and gain/ feed ratio were equivalent to that obtained with a mixture of soybean meal and methionine. This was the only meal that produced performance equivalent to that obtained with soybean meal and methionine. SUMMARY
ACKNOWLEDGMENTS
The commercial lysine supplement used in many of these experiments was provided by Merck and Co., Rahway, N.J. The antibiotic supplement was provided by Commercial Solvents Corporation, New York, N.Y. We are indebted to Mr. Harold M. Neilsen of the Chemistry Department, Utah State University, for the amino acid analysis of the cottonseed meal. REFERENCES Almquist, H. J., 1962. Amino Acid Composition of Poultry Feeds. Table published by Pacific Vegetable Oil Company, San Francisco, Cal. Anderson, J. O., and R. E. Warnick, 1965. Amino acid deficiencies in peanut meal and in corn and peanut meal rations. Poultry Sci. 44: 10661072. Dobson, D. C , J. O. Anderson and R. E. Warnick, 1964. A determination of the essential amino acid proportions needed to allow rapid growth in chicks. J. Nutrition, 82: 67-75. Grau, C. R., 1946. Protein concentrates as amino acid sources for the chick: corn gluten meal, cottonseed meal and peanut meal. J. Nutrition, 32: 303-312. Jonston, C , and A. B. Watts, 1964. The chick feeding value of meals prepared from glandless cottonseed. Poultry Sci. 43: 957-963. Kuiken, K. A., 1952. Availability of the essential amino acids in cottonseed meal. J. Nutrition, 46: 13-26. Snedecor, G. W., 1956. Statistical Methods, 5th Ed. The Iowa State College Press, Ames, Iowa. Spackman, D. H., W. H. Stein and S. Moore, 1958. Automatic recording apparatus for use in the chromatography of amino acids. Anal. Chem. 30: 1190-1206.
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Chicks were fed semi-purified rations containing 18 percent protein from a cottonseed meal and a mixture of the six strictly essential amino acids judged most apt to be deficient in the meal. Each of the strictly essential amino acids was replaced in turn with glycine to determine the sequence in which the essential amino acids become limiting in the cottonseed meal protein. Three commercial cottonseed meals and meals prepared in the laboratory from glanded and glandless cottonseed were used. Lysine was found to be the most limiting amino acid in the commercial meals and in the meal prepared in the laboratory from glanded seed. Methionine, isoleucine, threonine, and leucine were also found to be limiting in the order named. Methionine and lysine were found to be approximately equally limiting in the meal prepared in the laboratory from glandless seed. This meal is a much better source of lysine than the meal prepared from glanded seed; it also appeared to be a better source of methionine, isoleucine, and threonine.
89
EXPERIMENTAL
Five different cottonseed meals were used in these experiments. One commercial 1
Approved as Utah Agricultural Experiment Station Journal Paper No. 489.
84
expeller meal and two commercial prepress solvent-extracted meals were included. Prepress solvent-extracted meal B contained a higher protein level and a lower crude fiber level than meal A. Two meals were prepared under similar conditions at the Cottonseed Products Research Laboratory, Texas A & M University, College Station, Texas.2 One was prepared from glanded seed and the other from glandless seed. The crude fiber, moisture, and protein (N X 6.2S) contents, as well as the solubility of the nitrogen in 0.02N NaOH were determined for each lot of meal. These values are given in Table 1. The composition of the basic rations fed is given in Table 2. All rations fed in these experiments contained 19.1 percent protein (N X 6.25). The cottonseed meal level was varied according to its protein content. In all rations based on a given meal, enough of that meal was used to provide 18 percent protein to the ration. All cottonseed meal rations contained an amino acid mixture that provided an additional 1.1 per2
These meals were provided through Mr. A. Cecil Wamble of the Cottonseed Products Research Laboratory, Texas A & M University, College Station, Texas; and Mr. Garlon A. Harper of the National Cottonseed Products Association, Inc., Memphis, Tenn. The latter reported that chemical analysis indicated that the glanded and glandless meals contain the following, respectively: 0.28 and 0.47 percent residual lipids; 3.5 and 4.1 grams of lysine per 16 grams of nitrogen; 0.08 and 0.01 percent free gossypol; and 1.18 and 0.01 percent total gossypol. The meals were produced by direct hexane extraction; a moderate amount of heat was used during processing.
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IMITED amounts of meal prepared ' from glanded cottonseed are used in poultry rations. It has generally been recognized, however, that the protein in that meal is too deficient in lysine for large amounts to be included in commercial broiler rations. Grau (1946) found that lysine is the most limiting amino acid in cottonseed meal protein; additional methionine was also needed to allow the most rapid growth rate. Further supplementation with threonine or tryptophan did not increase growth rate. Jonston and Watts (1964) have reported that lysine and methionine are also limiting in meals prepared from glandless cottonseed. Lysine was found to be more limiting in the glanded than in the glandless meal. This paper reports the results of experiments conducted to determine the sequence in which the essential amino acids become limiting for growth of chicks in semipurified rations based on five different cottonseed meals. The experiments are similar to others recently reported with peanut meal (Anderson and Warnick, 196S). Our calculations indicated that a ration with 18 percent cottonseed meal protein would be deficient in amino acids other than lysine and methionine. These predictions were tested in these experiments.
L
AMINO ACID DEFICIENCIES IN COTTONSEED
85
TABLE 1.—Description of cottonseed meals fed
Type of cottonseed meal
Lot no.
Crude fiber Dry matter
("ptfvfiT'fl
Percent of ' air-dry sample 93 40.1
N soluble in 0.02N NaOH Percent 51
1
15.7
1 2
15.2 16.1
92 91
42.0 39.8
71 70
1 2
10.5 8.4
93 92
49.4 52.0
76 73
Prepared at Texas A & M from glanded seed
1
5.4
94
58.0
77
Prepared at Texas A & M from glandless seed
1 2
5.7 6.2
94 93
61.6 59.1
93 91
Expeller Prepress solvent-extracted Source A Prepress solvent-extracted Source B
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TABLE 2.—Composition of the basic rations cent protein. A ration based on soybean meal and methionine was also fed in all Cottonseed Soybean Ingredient experiments except one. The soybean meal meal meal used contained 50 percent protein and 4.0 Gms./kg. percent crude fiber. Cottonseed meal 295-452! — — 380 A sample of the second lot of prepress Soybean meal, 50% protein Corn oil 10 10 solvent-extracted meal B was hydrolyzed Salt 5 5 with 6N HCL for 24 hours. The levels of Dicalcium phosphate, 15 IS _18.5%P certain amino acids in the hydrolysate Limestone flour 10 10 2 0.6 0.6 were determined by the method of Spack- Trace mineral mix 3 1.4 1.4 Antibiotic supplement man et al. (1958). These levels were used Vitamin mix4 80 80 5 — L-lysine HC1 to estimate cottonseed protein content of DL-methionine 1 2 most amino acids as given in Table 3. The DL-isoleucine 1.5 — 2 — tryptophan and cystine estimates were DL-threonine 1 — DL-valine taken from a report by Almquist (1962). L-leucine 1 — 1 — The calculated level of each amino acid Glycine 1,000 1,000 Glucose monohydrate 5 to provided by a ration with 18 percent cot1 The cottonseed meal level varied with the tonseed protein and the level of each essenmeal's protein content; enough meal was used to tial amino acid in the ration found by provide 18 percent protein in the ration. 2 This mix provided the following in gms./kg. Dobson et al. (1964) to have a well balof ration: FeCeHsO,, 0.2; M n S 0 4 - H A 0.25; KI0 3 , anced mixture of the essential amino acids 0.01; ZnCOs, 0.13; and CuS0 4 , 0.01. 3 Provided 31 mgs. of zinc bacitracin/kg. of are also given in Table 3. The latter two ration. 4 sets of values were compared to predict Composition of this mix is given by Dobson which amino acids should be most limiting et al. (1964), This mix was designed to be used in purified rations at a 10 percent level. 6 in a ration with 18 percent cottonseed meal Cerelose, Corn Products Company, New York, N. Y. protein. When this series of experiments was calculations were based on Almquist's started, the amino acid analysis of cotton- (1962) values. When either set of values seed meal had not been done. The original was used, it was predicted that the same
86
J. 0. ANDERSON AND R. E. WAENICK
TABLE 3.—Essential amino acid content of cottonseed meal, a ration with 18 percent cottonseed protein, and the balanced ration of Dobson et a]. (1964)
Amino Acid
Gms.116 gms. N 4.0 1.5 3.51 3.4 3.6 6.3 5.0 2.7 1.3 1 5.5 8.3 11.5 4.4 21.0 9.0 3.8 3.9 4.4
Percent of ration 0.72 1.15 0.27 0.38 0.63 0.61 0.65 1.13 0.90 0.49 0.23 0.99
0.73 0.80 0.78 1.30 0.95 0.43 0.20
1.49 2.07 0.79 3.78 1.62 0.68 0.70 0.79
1.33 1.28
1 Values for cystine and tryptophan were taken from Almquist's (1962) table.
amino acids would be limiting in a ration with 18 percent cottonseed meal protein. The amino acid mixture added to the basic cottonseed meal ration fed in most experiments contained the six strictly essential amino acids predicted to be most limiting in a ration with 18 percent cottonseed protein. This mixture is termed the "complete" mixture in this paper. Valine was not included in the amino acid mixture of the basic ration fed in the first two experiments conducted with the expeller cottonseed meal. Glycine was added to all rations based on cottonseed meal. In each experiment with a given cottonseed meal, the basic ration and five or six other rations based on the same lot of cottonseed meal were fed. Each non-basic ration had one of the strictly essential amino acids removed from the complete amino acid mixture. More glycine was added to
RESULTS AND DISCUSSION
The results obtained are presented in Table 4. Lysine was found to be the most limiting amino acid in all of the commercial cottonseed meals and in the meal prepared in the laboratory from glanded seed. This was expected and comparable results
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Lysine Methionine Methionine & cystine Isoleucine Threonine Leucine Valine Histidine Tryptophan Phenylalanine Phenylalanine & Tyrosine Arginine Glycine Glutamic acid Aspartic acid Alanine Proline Serine
Level provided by Cottonseed 18% meal Cottonseeci Balanced content protein ration ration
these rations to keep the nitrogen level constant. A commercial lysine supplement containing 50 percent lysine was used in some experiments instead of the lysine hydrochloride. Results were similar when either source of lysine was used and have been summarized without regard to the lysine source. Chicks of both sexes from broiler crosses were used in all experiments. They were fed a practical-type ration from two to six days before the experimental rations were fed and were alloted to experimental groups on the basis of body weight just before being started on the experimental rations. Five birds composed each group. The experiments lasted from 14 to 21 days. All groups in an experiment started on experimental rations on the same date and ended on the same date. The birds were housed in electrically-heated, thermostatically controlled battery brooders with raised wire screen floors. Feed and water were supplied ad libitum. At the end of each experiment, the mean weight gain of each group of chicks was expressed as a percentage of the mean weight gain obtained with the basic cottonseed meal ration fed in that experiment. A relative gain/feed ratio was calculated in the same manner. All of the relative values obtained with any one ration were averaged to obtain the relative gains or gain/ feed ratios presented in Table 4. The relative group gains or gain/feed ratios were analyzed by the analysis of variance (Snedecor, 1956).
AMINO ACID DEFICIENCIES IN COTTONSEED
TABLE 4.—Results obtained with five cottonseed meals Amino acid removed from basic ration
Relative gain
Relative gain/feed ratio
Total no. groups
Expeller processed meal—3 experiments 8 100 100 None 1 6 27* 58* Lysine 6 78* 89* Methionine 8 85* 94* Isoleucine 8 91* 98 Threonine 3 92 94 Valine 8 99 101 Leucine Soybean meal & 2 5 99 methionine 114* Prepress solvent-extracted meal A—5 experiments 100 14 None 100 63* 11 Lysine 34* 88* 11 Methionine 70* 91* 13 Isoleucine 79* 92* 14 Threonine 84* 99 14 Leucine 95* 100 11 Valine 100 Soybean meal & 2 117* 13 methionine 99 Prepress solvent-extracted meal B—5 experiments 100 10 None 100 71* 10 Lysine 43* 83* 10 Methionine 68* 92* 10 Isoleucine 85* 96* 10 Threonine 90* 101 10 Leucine 97 99 10 Valine 99 Soybean meal & 109* 10 methionine2 103 Meal prepared from: glanded seed at Texas A 6* M— 2 experiments 100 100 None 4 44* 69* Lysine 4 65* 81* Methionine 4 73* 90* Isoleucine 4 79* 87* Threonine 4 93 97 Leucine 4 103 100 Valine 4 Soybean meal & 2 110* 110* methionine 4 Meal prepared from glandless seed at Texas A &° M—• 3 experiments 100 6 None 100 5 Methionine 73* 87* Lysine 73* 88* 5 93 4 Isoleucine 92 99 4 Leucine 93 4 Threonine 95 99 5 Valine 100 97 Soybean meal & 2 6 methionine 96 101 1 The basic ration fed in the first two experiments of this series did not include added valine. 2 This line gives the results obtained when the soybean meal ration (Table 2) was fed in the same experiments that the cottonseed meal was listed. * Significantly different than gain or gain/feed ratio obtained with cottonseed ration containing the complete amino acid mixture; P<0.01.
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have previously been reported by Grau (1946) who also conducted somewhat similar experiments with semi-purified rations. Many others have found lysine to be the most limiting amino acid in practicaltype rations based on cottonseed meal. The gains obtained when lysine was removed varied from 27 to 44 percent and the gain/ feed ratios varied from 58 to 71 percent of that obtained with the complete amino acid mixture. This suggests that the different meals prepared from glanded seed provided different amounts of available lysine to the chicks. Some of these differences may be due to the random variations in performance noted with this type of test. The low growth rate obtained when lysine was removed from the basic cottonseed meal rations raised a question as to whether the lysine level in the basic rations was adequate to allow' maximum growth rate. In one experiment with the second lot of prepress solvent-extracted meal A, an extra gram of lysine hydrochloride was added per kilogram of the basic ration in one treatment. This increase in lysine did not increase either growth rate or gain/ feed ratio. When lysine was omitted from basic rations using glanded cottonseed meals, the resultant amino acid imbalance probably decreased growth rate. In one experiment all of the essential amino acids were removed from the basic ration. Chicks fed this ration gained more than the chicks fed the ration from which only the lysine was removed from the complete amino acid mixture. Methionine was found to be the second limiting amino acid in all four meals prepared from glanded seed. Grau (1946) also found that methionine is limiting in cottonseed meal protein. Almquist (1962) indicates that cottonseed meal protein contains two grams of cystine per 16 grams of nitrogen. If this is true, then methionine and not the total level of sulphur-contain-
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ing amino acids is the limiting factor here. soybean meal ration. The ration made Isoleucine was found to be the third lim- from the meal prepared in the laboratory iting amino acid and threonine the fourth from glanded seed had about the same in each of the meals prepared from glanded crude fiber content as the ration based on seed. Leucine appears to be the fifth lim- soybean meal. The soybean meal ration iting in these meals although the reduction produced a significantly greater gain and in gain noted when leucine was removed better feed efficiency than the basic ration was only statistically significant with prep- made with the laboratory prepared meal. ress solvent-extracted meal A. The overall The meal prepared from glandless cotdifference in gain obtained when leucine tonseed is a much better source of lysine was removed from all four basic rations than any of the four meals prepared from was also statistically significant. glanded cottonseed. Methionine and lysine Valine removal from the different basic appeared to be about equally limiting in rations produced a marked reduction in the meal prepared from glandless cottonperformance only in the third experiment seed. Removal of isoleucine, threonine, or of the experiments conducted with the ex- leucine from this basic ration reduced peller processed meal. Valine was not in- growth rate and gain/feed ratio, but these cluded in the basic ration fed in the first differences were not statistically significant two experiments with this meal. The in these experiments. When lysine, methiodifference noted with valine removal in the nine, isoleucine, or threonine was removed third experiment probably is not real. The from the basic rations containing the two growth rate obtained with the basic ration laboratory prepared meals, the glandless without added valine in the second experi- meal produced the highest relative performent was equal to that obtained with the mance. soybean meal ration. The soybean meal raOne of the experiments with the glandtion was not fed in the first experiment. Our less meal and one with the glanded meal supply of this meal was exhausted after were done at the same time with chicks three experiments, and it was not consid- from the same hatch. Chicks fed the basic ered worthwhile to obtain a new lot for ad- ration with glandless cottonseed meal ditional tests. gained 16.8 grams per day with a gain/ The growth rates obtained with the feed ratio of 0.56, while chicks fed the basic rations made with commercial cot- basic ration with the glanded meal gained tonseed meals were comparable to those 14.5 grams per day with a gain/feed ratio obtained with the ration based on soybean of 0.51. Thus, the differences in absolute meal and methionine, but the soybean meal growth rate and gain/feed ratio were even ration produced significantly better feed greater than the relative differences given efficiency. The rations based on commercial in Table 4. The protein of the meal precottonseed meals contained from 1.5 to 5.8 pared from the glandless seed appears to percent more crude fiber than the ration provide more of at least these four amino based on soybean meal. All of these rations acids than the glanded meal protein. Kuikcontained a rather wide energy/protein en (1952) found that lysine availability ratio. The higher fiber level in the cotton- was greatly reduced by heating cottonseed seed rations probably made the growth meal with gossypol dissolved in cottonseed rate obtained with these rations higher oil. Luecine, isoleucine, valine, threonine, than would have been obtained if the crude and methonine availabilities were also refiber level had equaled that found in the duced by this treatment.
AMINO ACID DEFICIENCIES IN COTTONSEED
When this mixture of seven amino acids was added to the glandless cottonseed meal, the resulting growth rate and gain/ feed ratio were equivalent to that obtained with a mixture of soybean meal and methionine. This was the only meal that produced performance equivalent to that obtained with soybean meal and methionine. SUMMARY
ACKNOWLEDGMENTS
The commercial lysine supplement used in many of these experiments was provided by Merck and Co., Rahway, N.J. The antibiotic supplement was provided by Commercial Solvents Corporation, New York, N.Y. We are indebted to Mr. Harold M. Neilsen of the Chemistry Department, Utah State University, for the amino acid analysis of the cottonseed meal. REFERENCES Almquist, H. J., 1962. Amino Acid Composition of Poultry Feeds. Table published by Pacific Vegetable Oil Company, San Francisco, Cal. Anderson, J. O., and R. E. Warnick, 1965. Amino acid deficiencies in peanut meal and in corn and peanut meal rations. Poultry Sci. 44: 10661072. Dobson, D. C , J. O. Anderson and R. E. Warnick, 1964. A determination of the essential amino acid proportions needed to allow rapid growth in chicks. J. Nutrition, 82: 67-75. Grau, C. R., 1946. Protein concentrates as amino acid sources for the chick: corn gluten meal, cottonseed meal and peanut meal. J. Nutrition, 32: 303-312. Jonston, C , and A. B. Watts, 1964. The chick feeding value of meals prepared from glandless cottonseed. Poultry Sci. 43: 957-963. Kuiken, K. A., 1952. Availability of the essential amino acids in cottonseed meal. J. Nutrition, 46: 13-26. Snedecor, G. W., 1956. Statistical Methods, 5th Ed. The Iowa State College Press, Ames, Iowa. Spackman, D. H., W. H. Stein and S. Moore, 1958. Automatic recording apparatus for use in the chromatography of amino acids. Anal. Chem. 30: 1190-1206.
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Chicks were fed semi-purified rations containing 18 percent protein from a cottonseed meal and a mixture of the six strictly essential amino acids judged most apt to be deficient in the meal. Each of the strictly essential amino acids was replaced in turn with glycine to determine the sequence in which the essential amino acids become limiting in the cottonseed meal protein. Three commercial cottonseed meals and meals prepared in the laboratory from glanded and glandless cottonseed were used. Lysine was found to be the most limiting amino acid in the commercial meals and in the meal prepared in the laboratory from glanded seed. Methionine, isoleucine, threonine, and leucine were also found to be limiting in the order named. Methionine and lysine were found to be approximately equally limiting in the meal prepared in the laboratory from glandless seed. This meal is a much better source of lysine than the meal prepared from glanded seed; it also appeared to be a better source of methionine, isoleucine, and threonine.
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