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The Amino Acid Excretion Values in Intact and Cecectomized Negative Control Roosters Used for Determining Metabolic Plus Endogenous Urinary Losses1
The Amino Acid Excretion Values in Intact and Cecectomized Negative Control Roosters Used for Determining Metabolic Plus Endogenous Urinary Losses1 J. W. KESSLER,2 T. H. NGUYEN,3 and O. P. THOMAS Department of Poultry Science, University of Maryland, College Park, Maryland 20742 (Received for publication December 5, 1980) ABSTRACT Intact and cecectomized adult male, Single Comb White Leghorns were fasted for 24 hr prior to the initiation of the collection period. Excreta were quantitatively collected from each of the unfed control roosters for a 24 to 48 hr fasting period. The amino acids excretion values (mg/bird/24 hr) were not significantly different (P>.05) between the intact and cecectomized roosters for histidine or methionine. However, lysine and glutamic acid were significantly different (P<.05) and all other reported amino acids were significantly different (P<.01). The results of this preliminary experiment indicate the possible role of the ceca in altering the amino acid excretion values. Additional research is required in the elucidation of cecal involvement, as well as the selection of a proper control for this method of analysis for amino acid availability. (Key words- amino acid excretion, ceca, bioassay, fasting, lysine, methionine) 1981 Poultry Science 60:1576-1577
INTRODUCTION T h e t r u e metabolizable energy (TME) bioassay for determining t h e energy c o n t e n t of feedingstuffs was published by Sibbald ( 1 9 7 6 ) . This m e t h o d differs from t h e conventional m e t h o d s for determining t h e energy value of ingredients in t h a t it makes a correction for metabolic fecal plus endogenous urinary energy excretion. Likuski and Dorrell ( 1 9 7 8 ) r e p o r t e d a p p a r e n t and corrected amino acid availability values for corn and soybean meal force-fed t o roosters utilizing t h e TME bioassay as outlined b y Sibbald ( 1 9 7 6 ) . A bioassay t h a t would measure b o t h TME and true available a m i n o acids was published b y Sibbald ( 1 9 7 9 a ) . Sibbald ( 1 9 7 9 b ) determined a p p a r e n t and t r u e available amino acid values for corn, oats, wheat, and barley. T h e a p p a r e n t a m i n o acid values varied with t h e level of ingredient input, b u t when these values were corrected for metabolic plus endogenous excretion, t h e ' resulting true available a m i n o acid values were relatively uniform. Muztar et al. ( 1 9 8 0 ) published t r u e
'Scientific Article No. A2905, Contribution No. 5960 of the Maryland Agricultural Experiment Station (Department of Poultry Science). 2 Present Address: Poultry Research & Marketing, Ralston Purina Company, Checkerboard Square, St. Louis, Missouri 63188. 'Present Address: GuyoMarc'H, Service Technique B.P. 234, 56-Vannes-France.
amino acid availability values for soybean meal and rapeseed meal p r o d u c t s as determined in t w o cooperating laboratories. T h e resultant data showed reasonably good agreement for t h e t r u e a m i n o acid availability values b e t w e e n t h e t w o labs. T h e work outlined in this report describes t h e a m i n o acid excretion p a t t e r n in intact and cecectomized negative control (unfed) birds. The negative control bird is used t o d e t e r m i n e t h e correction factor for metabolic fecal and e n d o g e n o u s urinary amino acid excretion. MATERIALS AND METHODS Intact and cecectomized adult male, Single C o m b White Leghorns were used in this experim e n t . T h e cecectomized roosters were allowed 3 m o t o recover from surgery prior to being used in t h e assay. T h e birds were individually housed in wire cages and were fasted for 24 hr prior t o t h e initiation of t h e 24-hr collection period. Excreta samples were collected quantitatively, frozen, freeze-dried, equilibrated with a t m o s p h e r i c moisture, weighed, and ground as outlined by Sibbald ( 1 9 7 6 ) . Samples of excreta were h y d r o l y z e d for 24 hr with 6 N HCl under a nitrogen a t m o s p h e r e . A m i n o acid values were determined with a T e c h n i c o n Auto-Analyzer TSM-1. Because t h e r e is only one 24-hr hydrolysis, t h e results for isoleucine and valine have been divided b y . 8 5 . T h e resultant data were analyzed by a single variance analysis (F-test) (Snedecor, 1 9 5 6 ) .
1576
1577
RESEARCH NOTE TABLE 1. Mean values for amino acids excreted by fasted intact and cecectomized control roosters (mg/bird/24 br) Intact Alanine Arginine Aspartic acid Glutamic acid Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Proline Serine Threonine Tyrosine Valine
1 6 . 1 5 A ± 1.01' 1 7 . 1 3 A ± 1.12 30.64 A ± 1.90 49.00 a ±2.31 13.17 a ± .65 13.18 A ± .80 2 0 . 5 2 A ± 1.30 23.60 a ± 1.72 5.51 a ± .34 1 1 . 3 7 A ± .58 2 1 . 1 9 A ± 1.06 2 4 . 2 0 A ± 1.31 2 0 . 6 9 A ± 1.15 1 3 . 0 3 A ± .70 1 8 . 3 9 A ± 1.15
(18) 2 (18) (18) (18) (17) (18) (18) (16) (18) (18) (18) (18) (18) (18) (18)
Cecectomized
Sibbald (1979b)
20.71 B ± .84(11) 22.02 B ± 1.25 (11) 38.67 B ± 1.45 (11) 57.21 b + 2.36 (11) 13.66 a ± .38(11) 16.99 B ± .68(11) 28.07 B ± 1.29 (11) 28.32 b ± 1.11 (11) 6.16 a ± .24(11) 17.23 B ± 1.88(11) 34.80 B ± 1.96 (11) 38.78 B ± 1.57(11) 32.79 B ± 1.53 (11) 17.39 B ± 1.05 (11) 26.92 B ± 1.19(11)
17.3 20.6 30.2 48.4 12.0 13.0 25.5 21.4 5.0 12.2 24.4 26.6 20.8 15.0 17.0
±1.8(6) ±1.9 (6) ± 3.0(6) ±4.3 (6) ± 1.6 (6) ± 1.2(6) ± 2.9(6) ± 2.6 (6) ± .6 (6) ± 1.3 (6) ± 1.8(6) ± 2.1 (6) ± 2.2 (6) + 1.4 (6) ± 1.8(6)
' ' ' Means across rows with a superscript are not significantly different (P>.05); a,b significantly different (P<.05); and A,B significantly different (P<.01). 1
Mean ± standard error of mean.
2
Number of observations.
RESULTS AND DISCUSSION
The amino acid excretion values (mg/bird/24 hr) for the intact and cecectomized negative control birds are presented in Table 1. The values for methionine and histidine were not significantly different (P>.05) between the intact and cecectomized roosters. However, the amino acid values for lysine and glutamic acid were significantly different (P<.05) between the two groups of birds. All other amino acids were significantly different (P<.01) between the two groups of birds. In all cases the average value for the cecectomized roosters was greater than the intact roosters for each amino acid. The excreted amino acid values for the intact control birds were similar to those reported by Sibbald (1979b). Subsequently, Sibbald (1980) reported that the concentration of amino acids excreted by negative control birds decreased with the duration of the starvation period. The author also stated that the amino acid concentrations excreted by the negative control birds were similar to the values reported in the (1979b) publication. The significant differences reported for all amino acids with the exception of histidine and methionine indicate the ceca in some manner
are affecting amino acid excretion patterns. The mechanism by which the ceca may be affecting amino acid excretion was not ascertained in this preliminary experiment. However, the resulting data reported in this paper do indicate the need for further research in this
REFERENCES Likuski, H.J.A., and H. G. Dorrell, 1978. A bioassay for rapid determination of amino acid availability values. Poultry Sci. 57:1658-1660. Mutzar, A. M., S. J. Slinger, H.J.A. Likuski, and H. G. Dorrell, 1980. True amino acid availability values for soybean meal and tower and candle rapeseed and rapeseed meals determined in two laboratories. Poultry Sci. 59:605-610. Sibbald, I. R., 1976. A bioassay for true metabolizable energy in feedingstuffs. Poultry Sci. 55:303-308. Sibbald, I. R., 1979a. A bioassay for available amino acids and true metabolizable energy in feedingstuffs. Poultry Sci. 58:668-673. Sibbald, I. R., 1979b. Bioavailable amino acids and true metabolizable energy of cereal grains. Poultry Sci. 58:934-939. Sibbald, I. R., 1980. The effects of dietary cellulose and sand on the combined metabolic plus endogenous energy and amino acid outputs of adult cockerels. Poultry Sci. 59:836-844. Snedecor, G. W., 1956. Statistical methods. The Iowa State University Press, Ames, IA.
INTRODUCTION T h e t r u e metabolizable energy (TME) bioassay for determining t h e energy c o n t e n t of feedingstuffs was published by Sibbald ( 1 9 7 6 ) . This m e t h o d differs from t h e conventional m e t h o d s for determining t h e energy value of ingredients in t h a t it makes a correction for metabolic fecal plus endogenous urinary energy excretion. Likuski and Dorrell ( 1 9 7 8 ) r e p o r t e d a p p a r e n t and corrected amino acid availability values for corn and soybean meal force-fed t o roosters utilizing t h e TME bioassay as outlined b y Sibbald ( 1 9 7 6 ) . A bioassay t h a t would measure b o t h TME and true available a m i n o acids was published b y Sibbald ( 1 9 7 9 a ) . Sibbald ( 1 9 7 9 b ) determined a p p a r e n t and t r u e available amino acid values for corn, oats, wheat, and barley. T h e a p p a r e n t a m i n o acid values varied with t h e level of ingredient input, b u t when these values were corrected for metabolic plus endogenous excretion, t h e ' resulting true available a m i n o acid values were relatively uniform. Muztar et al. ( 1 9 8 0 ) published t r u e
'Scientific Article No. A2905, Contribution No. 5960 of the Maryland Agricultural Experiment Station (Department of Poultry Science). 2 Present Address: Poultry Research & Marketing, Ralston Purina Company, Checkerboard Square, St. Louis, Missouri 63188. 'Present Address: GuyoMarc'H, Service Technique B.P. 234, 56-Vannes-France.
amino acid availability values for soybean meal and rapeseed meal p r o d u c t s as determined in t w o cooperating laboratories. T h e resultant data showed reasonably good agreement for t h e t r u e a m i n o acid availability values b e t w e e n t h e t w o labs. T h e work outlined in this report describes t h e a m i n o acid excretion p a t t e r n in intact and cecectomized negative control (unfed) birds. The negative control bird is used t o d e t e r m i n e t h e correction factor for metabolic fecal and e n d o g e n o u s urinary amino acid excretion. MATERIALS AND METHODS Intact and cecectomized adult male, Single C o m b White Leghorns were used in this experim e n t . T h e cecectomized roosters were allowed 3 m o t o recover from surgery prior to being used in t h e assay. T h e birds were individually housed in wire cages and were fasted for 24 hr prior t o t h e initiation of t h e 24-hr collection period. Excreta samples were collected quantitatively, frozen, freeze-dried, equilibrated with a t m o s p h e r i c moisture, weighed, and ground as outlined by Sibbald ( 1 9 7 6 ) . Samples of excreta were h y d r o l y z e d for 24 hr with 6 N HCl under a nitrogen a t m o s p h e r e . A m i n o acid values were determined with a T e c h n i c o n Auto-Analyzer TSM-1. Because t h e r e is only one 24-hr hydrolysis, t h e results for isoleucine and valine have been divided b y . 8 5 . T h e resultant data were analyzed by a single variance analysis (F-test) (Snedecor, 1 9 5 6 ) .
1576
1577
RESEARCH NOTE TABLE 1. Mean values for amino acids excreted by fasted intact and cecectomized control roosters (mg/bird/24 br) Intact Alanine Arginine Aspartic acid Glutamic acid Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Proline Serine Threonine Tyrosine Valine
1 6 . 1 5 A ± 1.01' 1 7 . 1 3 A ± 1.12 30.64 A ± 1.90 49.00 a ±2.31 13.17 a ± .65 13.18 A ± .80 2 0 . 5 2 A ± 1.30 23.60 a ± 1.72 5.51 a ± .34 1 1 . 3 7 A ± .58 2 1 . 1 9 A ± 1.06 2 4 . 2 0 A ± 1.31 2 0 . 6 9 A ± 1.15 1 3 . 0 3 A ± .70 1 8 . 3 9 A ± 1.15
(18) 2 (18) (18) (18) (17) (18) (18) (16) (18) (18) (18) (18) (18) (18) (18)
Cecectomized
Sibbald (1979b)
20.71 B ± .84(11) 22.02 B ± 1.25 (11) 38.67 B ± 1.45 (11) 57.21 b + 2.36 (11) 13.66 a ± .38(11) 16.99 B ± .68(11) 28.07 B ± 1.29 (11) 28.32 b ± 1.11 (11) 6.16 a ± .24(11) 17.23 B ± 1.88(11) 34.80 B ± 1.96 (11) 38.78 B ± 1.57(11) 32.79 B ± 1.53 (11) 17.39 B ± 1.05 (11) 26.92 B ± 1.19(11)
17.3 20.6 30.2 48.4 12.0 13.0 25.5 21.4 5.0 12.2 24.4 26.6 20.8 15.0 17.0
±1.8(6) ±1.9 (6) ± 3.0(6) ±4.3 (6) ± 1.6 (6) ± 1.2(6) ± 2.9(6) ± 2.6 (6) ± .6 (6) ± 1.3 (6) ± 1.8(6) ± 2.1 (6) ± 2.2 (6) + 1.4 (6) ± 1.8(6)
' ' ' Means across rows with a superscript are not significantly different (P>.05); a,b significantly different (P<.05); and A,B significantly different (P<.01). 1
Mean ± standard error of mean.
2
Number of observations.
RESULTS AND DISCUSSION
The amino acid excretion values (mg/bird/24 hr) for the intact and cecectomized negative control birds are presented in Table 1. The values for methionine and histidine were not significantly different (P>.05) between the intact and cecectomized roosters. However, the amino acid values for lysine and glutamic acid were significantly different (P<.05) between the two groups of birds. All other amino acids were significantly different (P<.01) between the two groups of birds. In all cases the average value for the cecectomized roosters was greater than the intact roosters for each amino acid. The excreted amino acid values for the intact control birds were similar to those reported by Sibbald (1979b). Subsequently, Sibbald (1980) reported that the concentration of amino acids excreted by negative control birds decreased with the duration of the starvation period. The author also stated that the amino acid concentrations excreted by the negative control birds were similar to the values reported in the (1979b) publication. The significant differences reported for all amino acids with the exception of histidine and methionine indicate the ceca in some manner
are affecting amino acid excretion patterns. The mechanism by which the ceca may be affecting amino acid excretion was not ascertained in this preliminary experiment. However, the resulting data reported in this paper do indicate the need for further research in this
REFERENCES Likuski, H.J.A., and H. G. Dorrell, 1978. A bioassay for rapid determination of amino acid availability values. Poultry Sci. 57:1658-1660. Mutzar, A. M., S. J. Slinger, H.J.A. Likuski, and H. G. Dorrell, 1980. True amino acid availability values for soybean meal and tower and candle rapeseed and rapeseed meals determined in two laboratories. Poultry Sci. 59:605-610. Sibbald, I. R., 1976. A bioassay for true metabolizable energy in feedingstuffs. Poultry Sci. 55:303-308. Sibbald, I. R., 1979a. A bioassay for available amino acids and true metabolizable energy in feedingstuffs. Poultry Sci. 58:668-673. Sibbald, I. R., 1979b. Bioavailable amino acids and true metabolizable energy of cereal grains. Poultry Sci. 58:934-939. Sibbald, I. R., 1980. The effects of dietary cellulose and sand on the combined metabolic plus endogenous energy and amino acid outputs of adult cockerels. Poultry Sci. 59:836-844. Snedecor, G. W., 1956. Statistical methods. The Iowa State University Press, Ames, IA.