This article was downloaded by: [East Carolina University] On: 17 July 2013, At: 17:55 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
British Poultry Science Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/cbps20
Effect of deficiencies of single essential amino acids on nitrogen and energy utilisation in chicks a
Jun‐Ichi Okumura & Setsutaka Mori
a
a
Laboratory of Animal Nutrition, Faculty of Agriculture, Nagoya University, Nagoya‐shi, 464, Japan Published online: 08 Nov 2007.
To cite this article: JunIchi Okumura & Setsutaka Mori (1979) Effect of deficiencies of single essential amino acids on nitrogen and energy utilisation in chicks, British Poultry Science, 20:4, 421-429 To link to this article: http://dx.doi.org/10.1080/00071667908416601
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/ terms-and-conditions
Br. Poult. Sci., 20: 421-429. 1979
Longman: printed in Great Britain
EFFECT OF DEFICIENCIES OF SINGLE ESSENTIAL AMINO ACIDS ON NITROGEN AND ENERGY UTILISATION IN CHICKS JUN-ICHI OKUMURA AND SETSUTAKA MORI Laboratory of Animal Nutrition, Faculty of Agriculture, Nagoya University, Nagoya-shi 464, Japan
Downloaded by [East Carolina University] at 17:55 17 July 2013
Received for publication 8th November 1978
1. Diets 50% deficient in single essential amino acids were fed to chicks from day 8 to day 18 after hatching to evaluate body-weight gain, food consumption, body composition, nitrogen (N) and energy utilisation. 2. Body-weight gain was reduced most severely by deficiency of isoleucine followed in decreasing order by threonine, arginine, valine, histidine, tryptophan, methionine plus cystine, phenylalanine plus tyrosine, leucine and lysine, and possible reasons for the differences are discussed. 3. Body-weight gain and food efficiency were highly correlated with food consumption but metabolisable energy value of diets was not affected by single essential amino acid deficiencies. 4. Generally N retention (N retained/N consumed) and energy retention (energy retained/energy consumed) reflected food consumption, except for a lower N retention by chicks fed on the methionine plus cystinedeficient diet and for a lower energy retention by chicks fed on the valine deficient diet. 5. The amino acid deficient in the diet was present at very low concentration in the blood plasma. INTRODUCTION
Since the original proposal of Mitchell and Block (1946), it has been generally accepted that the nutritive quality of dietary protein can be predicted by calculating the percentage deficit of the most limiting amino acid, using the requirement pattern of essential amino acids as a standard. However, chick diets formulated to be equally limiting in different amino acids are not necessarily equally limiting in terms of ability to promote growth. Huston and Scott (1968) reported that chicks grow faster when fed on a diet deficient in leucine or arginine than when fed on a diet equally deficient in isoleucine. Sugahara et al. (1969) reported that deficiencies of phenylalanine plus tyrosine, tryptophan or isoleucine impaired growth more than equal deficiencies of other amino acids. The present study was undertaken to examine the effect on performance and body composition of feeding diets to chicks in which each of the classical 10 essential amino acids was decreased in turn to half of the amount considered to meet the requirements by Blair et al. (1972). 421
422
JUN-ICHI OKUMURA AND SETSUTAKA MORI
Downloaded by [East Carolina University] at 17:55 17 July 2013
MATERIALS AND METHODS
One-d-old, single comb, White Leghorn male chicks were housed in electrically-heated brooders. They received a proprietary chick mash for 7 d. On day 8 the chicks were individually weighed after starving for 2 h and were then selected and distributed into 12 groups of five chicks each, so that mean body weights were as uniform as possible. At this stage they were housed individually in metabolism cages (Okumura et al., 1978) for measurement of food consumption and quantitative collection of droppings. Eleven groups of five chicks each were fed on the appropriate experimental diets ad libitum from day 8 to day 18, while the five chicks of the last group were killed with diethyl ether and used to determine the initial body composition. Table 1 shows the composition of the standard diet which was designed to supply all the essential amino acids in amounts considered adequate by Blair et al. (1972). It also contained 10 g L-proline (Lee and Blair, 1972) and 100 g L-glutamic acid/kg. In the amino acid-deficient diets, the quantity of one essential amino acid was decreased to half of that in the standard diet, keeping the dietary nitrogen TABLE 1 Composition (g/fcg) of standard diet
Amino acid mixture1 Vitamin mixture2 Mineral mixture3 Corn oil Sucrose Cellulose Antacid mixture Choline chloride Corn starch
201-8 2-0
56-3 300
200-0 50-0 100 1-5
to 1 000
1 Composition ofamino acid mixture (g/kg diet): L-arginine. HC112-1, L-histidine.HCl 3-6, L-lysine. HC1 14-1, L-tyrosine 6-0, L-phenylalanine 6-0, L-tryptophan 1-5, L-methionine 4-5, L-cystine 3-5, L-threonine 5-5, L-leucine 12-0, L-isoleucine 5-0, L-valine 8-0, glycine 10-0, L-proline 10'0, L-glutamic acid 100-0. 2 Okumura et al. (1978). 3 Nesheim et at. (1962).
content constant by changing the amount of glutamic acid. On alternate days, the chicks were weighed and food consumption was measured. The droppings from each chick were collected into 200 ml 0-05M-sulphuric acid in deep, stainlesssteel trays beneath the metabolism cages. The acid prevented further microbial action in the droppings, and loss of ammonia. Droppings collected during the last 4 d were air-dried at 70 °C. Blood was taken by cardiac puncture at the end of the 10-d experimental period, transferred to heparin-treated centrifuge tubes and immediately centrifuged to separate the plasma. Equal volumes of plasma from the five chicks within each treatment were pooled and deproteinised with 5 volumes of picric acid solution (10 g/1) and stored —20 CC. After thawing, the samples were centrifuged, and the picric acid removed by passing the samples through a 20 x 80mm column of Dowex 50 in HC1. The samples were then dried by rotary evaporation at 45 °C and redissolved in sodium citrate buffer, pH 2-2. Amino acid analysis
Downloaded by [East Carolina University] at 17:55 17 July 2013
SINGLE AMINO ACID DEFICIENCIES
423
was performed by the method of Moore et al. (1958) using automatic ion-exchange liquid chromatography (JCL-3BC, JEOL Co., Tokyo). Tryptophan and cystine were not analysed. At the end of the experiment, all chicks were killed for carcass analysis. Carcasses were deep frozen and minced with a meat grinder. The minced carcasses were deep frozen again with solid carbon dioxide and minced again. This procedure was repeated twice and the mince was then dried at 55 °C for 48 h. Nitrogen (N) in the diets and carcasses was determined by a Kjeldahl procedure and carcass protein was defined as N x 6-25. Carcass fat was extracted overnight (about 16 h) with diethyl ether using a Soxhlet apparatus and determined gravimetrically. Gains in protein, fat and energy during the experimental period were estimated by subtracting the initial values of body composition from the final ones (day 18). Mean initial body weight of the chicks on day 8 was 71-2 g and they contained 8-1 and 16-2 g of fat and protein, respectively. Combustible energy of the diets, droppings and carcasses was measured with an automatic bomb calorimeter (Shimazu CA-3, Shimazu Co., Kyoto). Energy gain was calculated using the values of 39-12 and 23-68 kj/g for fat and protein, respectively. Metabolisable energy values, uncorrected for N retention, were estimated as described by Hill and Anderson (1958) using the total collection method. Heat production was estimated by subtracting retained energy in the body from metabolisable energy of the diet. Data were analysed statistically by Tukey's method (Yoshida, 1975). RESULTS
Table 2 shows the performance of chicks fed on diets deficient in a single essential amino acid in descending order of weight gains. Striking differences were observed not only in weight gain but also in food consumption and efficiency of food conversion. A deficiency of methionine plus cystine, tryptophan, histidine, valine, arginine, threonine or isoleucine resulted in a significant growth depression when compared with the standard diet. Food consumption and conversion efficiency TABLE 2 Performance of chicks fed on diets deficient in single essential amino acids, and metabolisable energy values of the diets (values are means of five birds for period 8 to 18 d old)
Deficient amino acid None Lys Leu Phe+Tyr Met+Cys Trp His Val Arg Thr He Pooled SE
Weight gain (g) 55-8 50-6 45-0 40-4 32-4* 31-2* 27-6* 16-0* 10-2* 6-0* -3-4* 3-9
Food consumption (g) 142-6 132-0 117-8 112-6 122-6 100-8* 104-2 90-2* 76-4* 73-0* 55-8* 6-9
Weight gain Food eaten 0-392 0-383 0-380 0-350 0-261 0-302 0-265 0-168* 0-121* 0-069* -0-082* 0-032
* Significantly different from that on the standard diet (P
British Poultry Science Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/cbps20
Effect of deficiencies of single essential amino acids on nitrogen and energy utilisation in chicks a
Jun‐Ichi Okumura & Setsutaka Mori
a
a
Laboratory of Animal Nutrition, Faculty of Agriculture, Nagoya University, Nagoya‐shi, 464, Japan Published online: 08 Nov 2007.
To cite this article: JunIchi Okumura & Setsutaka Mori (1979) Effect of deficiencies of single essential amino acids on nitrogen and energy utilisation in chicks, British Poultry Science, 20:4, 421-429 To link to this article: http://dx.doi.org/10.1080/00071667908416601
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/ terms-and-conditions
Br. Poult. Sci., 20: 421-429. 1979
Longman: printed in Great Britain
EFFECT OF DEFICIENCIES OF SINGLE ESSENTIAL AMINO ACIDS ON NITROGEN AND ENERGY UTILISATION IN CHICKS JUN-ICHI OKUMURA AND SETSUTAKA MORI Laboratory of Animal Nutrition, Faculty of Agriculture, Nagoya University, Nagoya-shi 464, Japan
Downloaded by [East Carolina University] at 17:55 17 July 2013
Received for publication 8th November 1978
1. Diets 50% deficient in single essential amino acids were fed to chicks from day 8 to day 18 after hatching to evaluate body-weight gain, food consumption, body composition, nitrogen (N) and energy utilisation. 2. Body-weight gain was reduced most severely by deficiency of isoleucine followed in decreasing order by threonine, arginine, valine, histidine, tryptophan, methionine plus cystine, phenylalanine plus tyrosine, leucine and lysine, and possible reasons for the differences are discussed. 3. Body-weight gain and food efficiency were highly correlated with food consumption but metabolisable energy value of diets was not affected by single essential amino acid deficiencies. 4. Generally N retention (N retained/N consumed) and energy retention (energy retained/energy consumed) reflected food consumption, except for a lower N retention by chicks fed on the methionine plus cystinedeficient diet and for a lower energy retention by chicks fed on the valine deficient diet. 5. The amino acid deficient in the diet was present at very low concentration in the blood plasma. INTRODUCTION
Since the original proposal of Mitchell and Block (1946), it has been generally accepted that the nutritive quality of dietary protein can be predicted by calculating the percentage deficit of the most limiting amino acid, using the requirement pattern of essential amino acids as a standard. However, chick diets formulated to be equally limiting in different amino acids are not necessarily equally limiting in terms of ability to promote growth. Huston and Scott (1968) reported that chicks grow faster when fed on a diet deficient in leucine or arginine than when fed on a diet equally deficient in isoleucine. Sugahara et al. (1969) reported that deficiencies of phenylalanine plus tyrosine, tryptophan or isoleucine impaired growth more than equal deficiencies of other amino acids. The present study was undertaken to examine the effect on performance and body composition of feeding diets to chicks in which each of the classical 10 essential amino acids was decreased in turn to half of the amount considered to meet the requirements by Blair et al. (1972). 421
422
JUN-ICHI OKUMURA AND SETSUTAKA MORI
Downloaded by [East Carolina University] at 17:55 17 July 2013
MATERIALS AND METHODS
One-d-old, single comb, White Leghorn male chicks were housed in electrically-heated brooders. They received a proprietary chick mash for 7 d. On day 8 the chicks were individually weighed after starving for 2 h and were then selected and distributed into 12 groups of five chicks each, so that mean body weights were as uniform as possible. At this stage they were housed individually in metabolism cages (Okumura et al., 1978) for measurement of food consumption and quantitative collection of droppings. Eleven groups of five chicks each were fed on the appropriate experimental diets ad libitum from day 8 to day 18, while the five chicks of the last group were killed with diethyl ether and used to determine the initial body composition. Table 1 shows the composition of the standard diet which was designed to supply all the essential amino acids in amounts considered adequate by Blair et al. (1972). It also contained 10 g L-proline (Lee and Blair, 1972) and 100 g L-glutamic acid/kg. In the amino acid-deficient diets, the quantity of one essential amino acid was decreased to half of that in the standard diet, keeping the dietary nitrogen TABLE 1 Composition (g/fcg) of standard diet
Amino acid mixture1 Vitamin mixture2 Mineral mixture3 Corn oil Sucrose Cellulose Antacid mixture Choline chloride Corn starch
201-8 2-0
56-3 300
200-0 50-0 100 1-5
to 1 000
1 Composition ofamino acid mixture (g/kg diet): L-arginine. HC112-1, L-histidine.HCl 3-6, L-lysine. HC1 14-1, L-tyrosine 6-0, L-phenylalanine 6-0, L-tryptophan 1-5, L-methionine 4-5, L-cystine 3-5, L-threonine 5-5, L-leucine 12-0, L-isoleucine 5-0, L-valine 8-0, glycine 10-0, L-proline 10'0, L-glutamic acid 100-0. 2 Okumura et al. (1978). 3 Nesheim et at. (1962).
content constant by changing the amount of glutamic acid. On alternate days, the chicks were weighed and food consumption was measured. The droppings from each chick were collected into 200 ml 0-05M-sulphuric acid in deep, stainlesssteel trays beneath the metabolism cages. The acid prevented further microbial action in the droppings, and loss of ammonia. Droppings collected during the last 4 d were air-dried at 70 °C. Blood was taken by cardiac puncture at the end of the 10-d experimental period, transferred to heparin-treated centrifuge tubes and immediately centrifuged to separate the plasma. Equal volumes of plasma from the five chicks within each treatment were pooled and deproteinised with 5 volumes of picric acid solution (10 g/1) and stored —20 CC. After thawing, the samples were centrifuged, and the picric acid removed by passing the samples through a 20 x 80mm column of Dowex 50 in HC1. The samples were then dried by rotary evaporation at 45 °C and redissolved in sodium citrate buffer, pH 2-2. Amino acid analysis
Downloaded by [East Carolina University] at 17:55 17 July 2013
SINGLE AMINO ACID DEFICIENCIES
423
was performed by the method of Moore et al. (1958) using automatic ion-exchange liquid chromatography (JCL-3BC, JEOL Co., Tokyo). Tryptophan and cystine were not analysed. At the end of the experiment, all chicks were killed for carcass analysis. Carcasses were deep frozen and minced with a meat grinder. The minced carcasses were deep frozen again with solid carbon dioxide and minced again. This procedure was repeated twice and the mince was then dried at 55 °C for 48 h. Nitrogen (N) in the diets and carcasses was determined by a Kjeldahl procedure and carcass protein was defined as N x 6-25. Carcass fat was extracted overnight (about 16 h) with diethyl ether using a Soxhlet apparatus and determined gravimetrically. Gains in protein, fat and energy during the experimental period were estimated by subtracting the initial values of body composition from the final ones (day 18). Mean initial body weight of the chicks on day 8 was 71-2 g and they contained 8-1 and 16-2 g of fat and protein, respectively. Combustible energy of the diets, droppings and carcasses was measured with an automatic bomb calorimeter (Shimazu CA-3, Shimazu Co., Kyoto). Energy gain was calculated using the values of 39-12 and 23-68 kj/g for fat and protein, respectively. Metabolisable energy values, uncorrected for N retention, were estimated as described by Hill and Anderson (1958) using the total collection method. Heat production was estimated by subtracting retained energy in the body from metabolisable energy of the diet. Data were analysed statistically by Tukey's method (Yoshida, 1975). RESULTS
Table 2 shows the performance of chicks fed on diets deficient in a single essential amino acid in descending order of weight gains. Striking differences were observed not only in weight gain but also in food consumption and efficiency of food conversion. A deficiency of methionine plus cystine, tryptophan, histidine, valine, arginine, threonine or isoleucine resulted in a significant growth depression when compared with the standard diet. Food consumption and conversion efficiency TABLE 2 Performance of chicks fed on diets deficient in single essential amino acids, and metabolisable energy values of the diets (values are means of five birds for period 8 to 18 d old)
Deficient amino acid None Lys Leu Phe+Tyr Met+Cys Trp His Val Arg Thr He Pooled SE
Weight gain (g) 55-8 50-6 45-0 40-4 32-4* 31-2* 27-6* 16-0* 10-2* 6-0* -3-4* 3-9
Food consumption (g) 142-6 132-0 117-8 112-6 122-6 100-8* 104-2 90-2* 76-4* 73-0* 55-8* 6-9
Weight gain Food eaten 0-392 0-383 0-380 0-350 0-261 0-302 0-265 0-168* 0-121* 0-069* -0-082* 0-032
* Significantly different from that on the standard diet (P
