The data reported herein are taken from a Master's thesis submitted to the Graduate Faculty, University of Missouri, 1958--P. [3].Digitized 2007 AES.Includes bibliographical references (page 8)

Pfander, W. H. (William Harvey), 1923-

Royer, Roy F.

Tribble, Leland Floyd, 1923-

University of Missouri: MOspace

RESEARCH BULLETIN 938 MAY, 1968 UNIVERSITY OF MISSOURI COLLEGE OF AGRICULTURE AGRICULTURAL EXPERIMENT STATION COLUMBIA, MISSOURI Changes in Free Amino Acid Levels in the Blood of Growing Swine as an Index to Protein Quality and Availability W. H. PFANDER, Roy F. ROYER, AND 1. F. TRIBBLE (Publication authorized May 7, 1968) COLUMBIA, MISSOURI SUMMARY Paper chromatography was used to study the free amino acid pattern in blood serum of four Hampshire gilts. 1. The free amino acids present in swine blood serum showed a characteristic pattern of increasing and decreasing at intervals after the pigs were fed a prac-tical type ration containing supplemental isoleucine. 2. The isoleucine-leucine content in the blood serum of Hampshire gilts fed an isoleucine supplemented ration increased at 3 hours after feeding, decreased at 6 hours, increased slightly again at 12 hours, and decreased markedly at 18 hours. 3. The approach described could aid in determining protein quality and avail-ability but is too laborious and inexact to be recommended for routine use. 4. Methionine appears to be the limiting amino acid in the ration used. Changes in Free Amino Acid Levels in the Blood of Growing Swine as an Index to Protein Quality and Availability W. H. PFANDER, Roy F. ROYER,' and L. F. Tribble2 INTRODUCTION The plasma amino concentration of several species has been used as an aid in the study of their protein utilization. Much of the work has been done with dogs (Howell, 1906; Van Slyke et al., 1912; Denton and Elvehjem, 1952; Dent and Schilling, 1948; Denton et al., 1953; Denton and Elvehjem, 1953). Informa-tion from the rat is available from the study of Gibson et al., 1951; on the chick from Charkey et at., 1953, and Richardson, 1953. One of the early studies with humans was that of Munto et at., 1952, which showed that glucose lowered the blood amino acids within one hour after its administration. Butter had no effect on blood amino acid composition. When this study was initiated, no work had been reported with pigs; therefore, the purpose of the experiments described herein was to see if similar methods could be used for examining the amino acid content of the blood of growing pigs and to determine if these techniques had promise for future studies. MATERIALS AND METHODS The two rations used in this study are shown in Table 1. They were identi-cal except for isoleucine content. Four Hampshire gilts weighing approximately 36 pounds were self-fed ra-tion 760 (Table 1) for two weeks. Each gilt was removed from its pen and fasted for 18 hours. A blood sample was taken and the pig was fed all of ration 760-A it would consume in 30 minutes. The pig was then placed in a collection crate and blood samples were taken at intervals of 3, 6, 12, and 18 hours after feeding. The samples taken while gilts were on ration 760 and after they had been fasted were obtained through a 16 gauge needle in the ear veins. When the pigs were in collection crates, blood from the pigs' tails was collected. Five milliliters of blood were collected into 15 milliliter centrifuge tubes. The blood was centri-fuged at 3000 rpm for thirty minutes. A protein-free filtrate was pr~pared by the method of Folin and Wu (1919), frozen, and stored in a freezer at -lOoF until ready for analysis by paper chroma-tography according to the follOWing method. 1 The data reported herein are taken from a Master's thesis submitted to the Graduate Faculty, University of Missouri, 1958. 2 Present address: Animal Husbandry Department, Texas Technological College , Lubbock, Texas. 6 MISSOURI AGRICULTURAL EXPERIMENT STATION line of developed spots designated as zero hours shows the free amino acid pat-rem in blood serum after an IS-hour fast. Other verticle lines of developed spots show the free amino acid pattern in blood serum of the same gilt at intervals after she was fed ration 760-A. The combined isoleucine-leucine content is shown by the horizontal line of developed spots designated 1; numbers 2, 3, and 4 cor-respond to spots developed for methionine, tryptophan, and lysine plus histidine, respectively. The relative isoleucine-leucine content in blood serum from each gilt is shown in Figure 2. Although considerable variation existed among the serum isoleucine-leucine contents of the four gilts, the general pattern, as shown by the mean values, was an increase at 3 hours and a decrease at 6 hours after feeding. The content of isoleucine-leucine in swine blood serum after an IS-hour fast was determined as 9.4 micrograms per milliliter. An analysis of blood samples taken three hours after the pigs were fed ration 760-A showed the isoleucine-leucine content as 19.0 micrograms per milliliter of blood serum. At 6, 12, and 18 hours after feeding pigs ration 760-A, the blood serum levels of isoleucine-leucine were 7.9, 10.8, and 6.5 micrograms per milliliter, respectively. The con-centrations of the other amino acids were not determined quantitatively, but it appears that the concentration of tryptophan and methionine are very low and that lysine was present in large amounts. FIGURE 2 0.07 f RELATIVE ISOLEUCINE - LEUCINE CONTENT OF BLOOD SERUM AfTER FEEDING RATION 760 AND 760 A \ 0.06 ~ I 0.04 0. 03 0.02 0. 01 30 Each initia l va lue ob teined 18 hrs. ofter last feed on ration 760. Subsequent values after feeding 760A. o Midnight 300 600 900 1200 (noon) 1500 TIME 1800 2100 2400 2700 3000 RESEARCH BULLETIN 938 7 The free amino acid pattern of swine blood serum at intervals after feeding pigs rations containing crystalline isoleurine is in agreement with the findings of Denton and Elvehjem (1953) on dogs. The results of this experiment suggest that the supplemental isoleucine was rapidly absorbed from the small intestines, resulting in an increase of free iso-leucine in the blood. As glucose and amino acids liberated from the ingested ration became available, the free isoleucine in the blood was lowered by the ana-bolic processes of the body. The slight increase in the isoleucine-leucine content, 12 hours after feeding ration 760-A, may have corresponded to a reduced glucose supply followed by hydrolysis of labile body proteins. The second decrease in the isoleucine-leucine content of swine blood serum may have been a result of a deficiency of glucose or reduced TPN to furnish energy for synthetic reactions and amino acids may have been deaminized. The isoleucine-leucine content of swine blood serum was greater 18 hours after feeding ration 760 than it was 18 hours after feeding ration 760-A. The gilts were fed ration 760 ad libitum for two weeks before the first 18-hour fast. One blood sample was taken at the end of the first fast and the gilts were fed all of ration 760-A they would consume in one-half hour. A second 18-hour fast was started immediately and during this fast four blood samples wer/e taken. It is logical to believe that the supply of free amino acids in the blood was at a lower level at the start of the second fasting period than at the start of the first fast. Also, the added stress placed on the gilts during the second period may have helped reduce the free amino acids in the blood. It would have been pref-erable to have obtained values at intervals on each ration. The differences in the serum isoleucine-leucine content of the four Hamp-shire gilts may have been due in part to a diurnal variation. At each interval, blood samples were taken at the same time of day from Hampshire gilts 31 and 61. Blood samples were taken from the other gilts at a different time of day for the various intervals. There was less variation in the blood serum isoleucine-leucine content of Hampshire gilts 31 and 61 than for the other gilts, except at the 12-hour interval. Qualitative examination of the chromatographs suggested that methionine was the limiting amino acid in this ration and that lysine was being made avail-able in amounts exceeding the pig's ability to use it. 8 MISSOURI AGRICULTURAL EXPERIMENT STATION BIBLIOGRAPHY Block, Richard J. 1952. "Color Development on Paper," Paper Chromatography, A Lab-oratory Manual, 58. Charkey, L. W., Wm. K. Manning, A. K. Kano, F. X. Gassner, M. L. Hopwood and 1. L. Madsen. 1953. "A Further Study of Vitamin B12 in Relation to Amino Acid Metabolism in the Chick," Poultry Sci., 32 :630. Dent, C. E. and J. A. Schilling. 1948. "Studies on the Absorption of Proteins: The Amino-Acid Pattern in the Portal Blood," Biochem. J., 44:318. Denton, A. E. and C. A. Elvehjem. 1952. "Enzymatic Liberation of Amino Acids from Different Proteins," J. Nutr., 49:221. Denton, A. E., S. N. Gershoff and C. A. Elvehjem, 1953a. "A New Method of Can-nulating the Portal Vein of Dogs," J. Bioi. Chem., 204:731. Denton, A. E. and C. A. Elvehjem. 1953b. '.'Amino Acid Concentration in the Portal Vein after Ingestion of Amino Acid," J. Bioi. Chem., 206:455. Folin, Otto and Hsien Wu. 1919. "A System of Blood Analysis," J. Bioi. Chem., 38:81. Gibson, Q. H. and G. Wiseman. 1951. "Selective Absorption of Stereo-isomers of Amino-acids from Loops of the Small Intestine of the Rat," Biochem. J., 48:426. Howell, W. H. 1906. "Note upon the Presence of Amino Acids in the Blood and Lymph as Determined by the ,8-Naphthalinsulphochloride Reaction," Am. J. Physiol., 17:273. Munro, H. N. and W. S. T. Thomson. 1952. "The Response of Blood Amino-acids in the Fasting Subject to Glucose and to Fat Administration," Abstracts of Com-munications Brit. J. Nutr., 6-Pii. Richardson, L. R., Lynn G. Blaylock and Carl M. Lyman. 1953. "Influence of Dietary Amino Acid Supplements on the Free Amino Acids in the Blood Plasma of Chicks," J. Nutr., 51:515. Slotta, K. H. and J. Primosigh. 1951. "Amino-acid Composition of Crotoxin," Nature, 168:69G. Van Slyke, Donald D. and Gustav M. Meyer. 1912. "The Amino-Acid Nitrogen of the Blood. Preliminary Experiments on Protein Assimilation," J. Bioi. Chem., 12:399. 

Changes in free amino acid levels in the blood of growing swine as an index to protein quality and availability

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