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The Effects of Fasting Upon Plasma Enzyme Levels in Chickens
994
B. F. MILLER, P. E. SANFORD AND R. E. CLEGG
REFERENCES Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-42. Gabuten, A. R., and C. S. Shaffner, 1954. A study of the physiological mechanisms affecting specific gravity of chicken eggs. Poultry Sci. 33 : 47-53. Taylor, L. W., and B. R. Burmester, 1940. Effect of thyroidectomy on production, quality and composition of eggs. Poultry Sci. 19: 326-331. Winchester, C. F., 1939. Influence of thyroid on egg production. Endocrin. 24: 697-701.
Winchester, C. F., 1940. Growth and development with special reference to domestic animals. II. Seasonal metabolic and endocrine rhythms in the domestic fowl. Missouri Agr. Expt. Sta. Res. Bull. 315: 1-52. Winchester, C. F., C. L. Comer and G. K. Davis, 1949. Thyroid destruction by I 1 ", and replacement therapy. Science, 110: 302-304. Winchester, C. F., and V. C. Scarborough. 1953. The influence of thyroxine on growth and egg production of chickens. Poultry Sci. 32: 423429.
Lois S. MCDANIEL AND HELEN A. DEMPSEY Department of Animal Pathology, University of Maine, Orono, Maine (Received for publication November 6, 1961)
A LTERATIONS in the levels of seven l \ tricarboxylic acid enzymes and citric acid in the livers of chicks infected with Salmonella pullorum were observed by Gilfillin et al. (1956). Hill et al. (1960) also detected elevated levels of citric acid blood of chicks infected with Salmonella gallinarum. But the citric acid levels of the infected chicks were comparable with those of the controls if feed was withheld from both groups for 18 hours. Furthermore, withdrawal of feed for 36 hours from normal chicks resulted in elevated blood and liver citric acid levels. These were interesting observations because anorexia is a common symptom in many diseases. Although several papers about the effects of diet upon enzyme activities in chickens have been published, little information relative to the effects of actual fasting upon enzyme activities in birds is available. Stutts et al. (1957), however, detected a decrease in the plasma alkaline phosphatase This investigation was supported by PHS research grant C-4957 from the National Institutes of Health, Public Health Service.
(alkPH) levels of adult White Leghorns. Starvation decreases the concentration of glycogen in the avian liver, but does not affect the concentration of glycogen in the glycogen body which is located on the dorsal aspect of the spinal cord (Szepsenwol and Michalski, 1951). Studies on the effects of lack of feed and/ or water upon plasma enzyme levels in chickens are presented in this paper. The activity levels of lactic dehydrogenase (LDH), malic dehydrogenase (MDH), glutamic oxalacetic transaminase (GOT), aldolase (ALD), and alkPH were measured. EXPERIMENTAL
The experimental birds were hatched and housed in isolation units. Single Comb White Leghorns, Connecticut random breds, and sex-link crosses between Rhode Island Reds and Barred Plymouth Rocks were used. They received a complete growing ration which is manufactured commercially. The chickens in each experiment were all of the same "breed" and from the same hatch. The males and females were as
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
The Effects of Fasting Upon Plasma Enzyme Levels in Chickens
995
PLASMA ENZYME LEVELS TABLE 1.—The effects of withdrawal of feed upon plasma enzyme levels in chickens
Age
Treatment
Plasma enzyme levels (mean±S.D.)
No. birds
LDH
MDH
GOT
ALD
alkPH
Fasted Control
11 10
1,202 + 192 1,777 + 483
No feed 24 hours 734 + 222 833 ±229
355 + 152 363 + 185
35.2 + 3.5 35.5±5.6
not done
9 wk.
Fasted Control
7 7
989 + 267 1,028 + 205
635 + 117 702 ±144
370+163 340 ±180
37.0 + 2.6 39.7±2.2
not done
13 wk.
Fasted Control
13 14
l,319+448f 847 + 221
1,076 ±304 852 ±288
221± 48 186± 38
18.1+6.2 23.4±5.7
not done
10 wk.
Fasted Control
16 17
885 + 259* 706 + 188
745 + 150 671 + 177
151± 29 156+ 34
18.5 + 4.8 16.2±5.6
72 + 60 119±90
16 wk.
Fasted Control
12 12
258 + 168 267± 99
35.8 + 4.8f 30.0±3.3
not done
10 wk.
Fasted Control
14 14
886 + 279f 549 + 147
710+140+ 403 ± 71
144+ 30* 118+ 31
28.5 + 5.7f 18.8 + 5.7
not done
10 wk.
Fasted Control
14 15
l,154±268t 826 + 228
l,232±325t 851 ±230
140+ 36 154± 45
30.4 + 3.Of 25.5±2.6
not done
10 wk.
Fasted Control
16 17
l,229+389t 706 ±188
973 +285 f 671 ±177
173+ 48 156± 34
20.4 + 6.4 16.2±5.6
59 ±28* 119±90
No feed 48 hours 1,139 ±249* l,848 + 588f 886 ±224 1,089±264
*P<.05. fP<.01. Plasma enzyme levels are expressed as enzyme units per ml. of plasma. One LDH, MDH, or GOT unit is equivalent to the amount of enzyme activity which will cause a decrease of 0.001 in optical density per minute under standard conditions. ALD activity is expressed in dihydroxyacetone units. AlkPH activity is expressed in King-Armstrong units per 100 ml.
nearly equally divided as possible between the fasted and control groups. In four of the experiments, one group of normal birds served as controls for more than one group of fasted birds. Heparinized blood samples were collected by cardiac puncture. The alkPH activity was assayed by a modification of the King-Armstrong technique (Powell and Smith, 1954) on an Auto-Analyzer. It was necessary to dilute the plasma samples one to five for this test. The ALD activity was measured by the method of Friedman and Lapan (1958) on a Klett Summerson photoelectric colorimeter. The LDH, MDH, and GOT assays were conducted on a Beckman DU spectrophotometer. The methods for measuring LDH and MDH levels were previously described (McDaniel and Chute, 1961). The
GOT activity was assayed by the method of Karmen (1955) as described in Sigma Technical Bulletin No. 410 (1958). In the latter test, 0.1 ml. of plasma and 400 units of malic dehydrogenase were used. The data were analyzed by means of the "t" test (Snedecor, 1956). RESULTS The effects of the withdrawal of feed upon the plasma enzyme levels are given in Table 1. The removal of feed for 24 hours did not affect the MDH, GOT, and ALD levels, slightly decreased the alkPH levels, and occasionally increased the LDH levels. The removal of feed for 48 hours, however, caused the LDH, MDH, and ALD levels to increase and the alkPH levels to decrease. The GOT levels were signifi-
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
8 wk.
996
L. S. MCDANIEL AND H. A.
DEMPSEY
TABLE 2.—The effects of withdrawal of water upon plasma enzyme levels in chickens Age
No. Treatment birds
Plasma enyzme levels (mean + S.D.) ALD
alkPH
No water 24 hours 472 + 108* 126 ± 3 4 371± 75 127 + 22
19.0±1.8f 15.2±3.2
not done
LDH
MDH
GOT
Fasted Control
11 11
542 ±157 452+ 86
16 wk.
Fasted Control
16 15
679 + 152 732 + 127
612 + 166f 407± 88
108 + 23 104 ±32
16.6+4.5* 13.6 + 2.4
not done
5wk.
Fasted Control
32 31
691 + 168f 1,165 ±270
954±154f 733 ±150
190+32 192 + 33
18.5±2.8t 16.0±2.6
141 + 101 113 ± 68
10 wk.
Fasted Control
11 11
164±32f 127 + 22
23.9±4.1f 15.2±3.2
not done
6 wk.
Fasted Control
23 22
864±235f 1,165 + 204
870 + 151 941 ±126
181+34* 156 ±29
19.1+3.6 20.1±2.9
203 ±164 223 + 123
5wk.
Fasted Control
33 32
980 + 263 1,034 + 199
901 + 164 904 ±155
191 ± 4 1 184 + 32
21.5±4.7* 17.8 + 5.0
136 +531 201±108
10 wk.
Fasted Control
12 11
169 + 33f 127 ±22
37.9 + 5.3f 15.2±3.2
not done
No water 48 hours 520 + 150 475 + 143* 371± 75 452+ 86
No water 72 hours 457 ±149 446 + 113 452+ 86 371± 75
*P<.05. fP<.01. Enzyme activity units same as used in Table 1.
cantly elevated in only one of the four experiments. The effects of lack of water upon the plasma enzyme levels are presented in Table 2. Withdrawal of water for 24 hours caused an increase in the MDH activity, but the levels of this enzyme returned to normal after 48 or 72 hours. In general, water deprivation caused the LDH levels to decrease and the ALD levels to increase. The alkPH levels were depressed and the GOT levels were elevated when the chickens were fasted for 48 hours. The data in Table 3 indicate that removal of both feed and water for periods up to 48 hours did not affect the LDH, MDH, or GOT levels. The fasting did cause elevated ALD levels and slightly depressed alkPH levels. These results were reversed when chickens were given access to feed and water for 24 hours after they had been deprived of feed and water for
48 hours. Elevated LDH, MDH, and GOT levels and normal ALD and alkPH levels were detected. No statistically significant differences were noted between the plasma enzyme levels of the males and females within the fasted and control groups. Since more than one "breed" was used in these experiments, statistical analysis of the relationship of age to tolerance to fasting was not considered valid although similar results were obtained with all of the different "breeds." DISCUSSION
The standard deviations listed in the tables indicate the considerable individual variation in plasma enzyme levels which exists among chickens. They also indicate an apparent individual variation in the ability of the avian metabolism to withstand fasting. These observations agree with those of Stutts et al. (19S7) on a non-in-
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
10 wk.
997
PLASMA ENZYME LEVELS TABLE 3.—The effects of withdrawal of feed and water upon plasma enzyme levels in chickens
Age
Treatment
No. birds
Plasma enzyme levels (mean±S.D.) LDH
MDH
GOT
ALB
alkPH
Fasted Control
11 12
No feed
22.3 + 3.3 21.2±3.3
not done
7wk.
Fasted Control
17 17
1,019 ±233 1,088 ±276
743 + 139 776 + 139
183 ± 34 191 ± 47
20.1±2.4 19.2±3.2
102± 47 135± 86
7wk.
Fasted Control
21 21
793 ±187 677 ±188
662 ±114 626 ±173
139 + 50 146 + 29
20.1±9.6f 13.2±4.9
96 ± 38 141 ± 83
7wk.
Fasted Control
17 17
18.9±3.2 19.2±3.2
105 ±123 135± 86
7wk.
Fasted Control
21 21
855 ±266 677 ±188
716 + 169 626 + 173
125 ± 39 146 ± 29
18.6±7.1f 13.2±4.9
80 + 22* 141 ± 83
7 wk.
Fasted Control
21 21
1,104 + 285 1,108 ±238
1,003 + 199 893 + 164
278 + 149 223+ 86
23.2 + 8.2f 13.9+4.4
97 ± 70 185 ±189
10 wk.
Fasted Control
10 wk.
Fasted Control
17 17
l,193±414f 752 ±221
877 + 257f 622 + 212
151 + 33* 125 + 30
24.5 + 4.6 20.7+4.6
93+ 37 81 ± 31
10 wk.
Fasted Control
28 30
1,103 + 249 1,014 + 222
801 + 170 830+139
136+14* 124+ 27
15.7 + 2.5 17.4 + 6.2
79 ± 31 85 ± 42
No feed
Access to feed and watei• for 24 hours after fasting 48 hours 837±217f 162+ 35 13.2 + 2.6 20 l,176 + 452t 20 662 + 113 557 + 118 152+ 32 11.6 + 1.7
90+ 38 95+ 55
*P<.05. fP<.01. Enzyme activity units same as used in Table 1.
bred strain of White Leghorns. These factors probably account for the apparent discrepancies in the data presented in this paper. Each enzyme studied appeared to follow an individual trend during fasting. Distinctive trends for enzymes have also been observed in rat livers during fasting (Soberon and Sanchez, 1961). It is interesting that the removal of both feed and water appeared to affect the plasma enzyme levels less than the removal of either feed or water alone. This was especially true in the case of the LDH, MDH, and GOT levels. Twenty-four hours appeared to be a sufficient length of time to allow the alkPH and ALD activities to return to the normal levels after the birds had fasted. On the
other hand, the sudden intake of nutrients appeared to disturb the LDH, MDH, and GOT levels more than the withdrawal of feed and water. If blood samples were collected after the birds were given access to feed and water for a longer period of time, the levels of these enzymes probably would have been normal. Since birds possess a rapid metabolism, it might be assumed that fasting would markedly affect blood enzyme levels. The results of these investigations disagree with this assumption. Even in the experiments where statistically significant differences in the average plasma enzyme levels were found, the ranges of the enzyme levels of the fasted birds and the controls overlaped. In physiologic or genetic studies, the effects
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
5wk.
998
L. S. MCDANIEL AND H. A. DEMPSEY
of fasting could be important factors. In pathologic studies, however, the enzyme levels of the affected chickens would have to be beyond the range of the enzyme levels of the normal birds to be considered significant. Therefore, anorexia probably would not appreciably affect the results of experiments on plasma enzyme levels in chickens during a pathologic condition.
Studies on the effects of the removal of feed and/or water upon plasma enzyme levels in chickens were conducted. During the three fasting conditions, the aldolase (ALD) levels were usually increased and the alkaline phosphatase (alkPH) levels were usually decreased. No statistically significant differences were noted in the alkPH levels until the birds had been fasted 48 hours. Lack of water affected the ALD levels more than the other two fasting conditions. The withholding of both feed and water for 48 hours did not affect the lactic dehydrogenase (LDH), malic dehydrogenase (MDH), and glutamic oxalacetic transaminase (GOT) levels. The LDH levels usually increased when feed was withheld and usually decreased when water was withheld. The MDH levels were not affected when feed was withheld for 24 hours, but were elevated when feed was withheld for 48 hours. Removal of water for 24 hours caused the MDH levels to increase, but the levels tended to return to normal after water had been removed for 48 hours or more. The GOT levels were elevated when water was withheld for 48 hours, but they were usually not affected when feed was withheld. When chickens were given access to feed
REFERENCES Friedman, M. M., and B. Lapan, 1958. Serum aldolase in the neonatal period: including a colorimetric determination of aldolase by standardization with dihydroxyacetone. J. Lab. Clin. Med. S i : 745-752. Gilfillan, R. F., D. F. Holtman and R. T. Ross, 1956. Influence of Salmonella pullorum infection on various liver tricarboxylic acid enzymes and citrate levels in the chick. J. Bact. 72: 624-627. Hill, C. H., H. W. Garren, J. W. Kelly and M. K. Warren, 1960. Studies on blood and liver citric acid levels in chicks. Poultry Sci. 39: 117-119. Karmen, A., 1955. A note on the spectrophotometric assay of glutamic oxalacetic transaminase in human blood serum. J. Clin. Invest. 34: 131-133. McDaniel, L. S., and H. L. Chute, 1961. Enzyme activity levels in chicken plasma. Am. J. Vet. Res. 22: 99-103. Powell, M. E. A., and M. J. H. Smith, 1954. The determination of serum acid and alkaline phosphatase activity with 4-aminoantipyrine (A.A.P.). J. Clin. Path. 7: 245-248. Sigma Technical Bulletin No. 410, 1958. Determination of glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase at 340mn in serum or other fluids. Sigma Chemical Co., St. Louis, Mo. Snedecor, G. W., 1956. Statistical Methods. 5th Ed. Iowa State College Press., Ames, la. Soberon, G., and E. Sanchez, 1961. Changes in effective enzyme concentration in the growing rat liver. I. Effects of fasting followed by repletion. J. Biol. Chem. 236: 1602-1606. Stutts, E. C , W. E. Briles and H. O. Kunkel, 1957. Plasma alkaline phosphatase activity in mature inbred chickens. Poultry Sci. 36: 269276. Szepsenwol, J., and J. V. Michalski, 1951. Glycogenolysis in the liver and glycogen body of the chicken after death. Am. J. Physiol. 165: 624-627.
AUGUST 9-15, 1963. SIXTH INTERNATIONAL CONGRESS ON NUTRITION, EDINBURGH, SCOTLAND
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
SUMMARY
and water for 24 hours after they had been denied both feed and water for 48 hours, elevated LDH, MDH, and GOT levels and normal ALD and alkPH levels were detected.
B. F. MILLER, P. E. SANFORD AND R. E. CLEGG
REFERENCES Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-42. Gabuten, A. R., and C. S. Shaffner, 1954. A study of the physiological mechanisms affecting specific gravity of chicken eggs. Poultry Sci. 33 : 47-53. Taylor, L. W., and B. R. Burmester, 1940. Effect of thyroidectomy on production, quality and composition of eggs. Poultry Sci. 19: 326-331. Winchester, C. F., 1939. Influence of thyroid on egg production. Endocrin. 24: 697-701.
Winchester, C. F., 1940. Growth and development with special reference to domestic animals. II. Seasonal metabolic and endocrine rhythms in the domestic fowl. Missouri Agr. Expt. Sta. Res. Bull. 315: 1-52. Winchester, C. F., C. L. Comer and G. K. Davis, 1949. Thyroid destruction by I 1 ", and replacement therapy. Science, 110: 302-304. Winchester, C. F., and V. C. Scarborough. 1953. The influence of thyroxine on growth and egg production of chickens. Poultry Sci. 32: 423429.
Lois S. MCDANIEL AND HELEN A. DEMPSEY Department of Animal Pathology, University of Maine, Orono, Maine (Received for publication November 6, 1961)
A LTERATIONS in the levels of seven l \ tricarboxylic acid enzymes and citric acid in the livers of chicks infected with Salmonella pullorum were observed by Gilfillin et al. (1956). Hill et al. (1960) also detected elevated levels of citric acid blood of chicks infected with Salmonella gallinarum. But the citric acid levels of the infected chicks were comparable with those of the controls if feed was withheld from both groups for 18 hours. Furthermore, withdrawal of feed for 36 hours from normal chicks resulted in elevated blood and liver citric acid levels. These were interesting observations because anorexia is a common symptom in many diseases. Although several papers about the effects of diet upon enzyme activities in chickens have been published, little information relative to the effects of actual fasting upon enzyme activities in birds is available. Stutts et al. (1957), however, detected a decrease in the plasma alkaline phosphatase This investigation was supported by PHS research grant C-4957 from the National Institutes of Health, Public Health Service.
(alkPH) levels of adult White Leghorns. Starvation decreases the concentration of glycogen in the avian liver, but does not affect the concentration of glycogen in the glycogen body which is located on the dorsal aspect of the spinal cord (Szepsenwol and Michalski, 1951). Studies on the effects of lack of feed and/ or water upon plasma enzyme levels in chickens are presented in this paper. The activity levels of lactic dehydrogenase (LDH), malic dehydrogenase (MDH), glutamic oxalacetic transaminase (GOT), aldolase (ALD), and alkPH were measured. EXPERIMENTAL
The experimental birds were hatched and housed in isolation units. Single Comb White Leghorns, Connecticut random breds, and sex-link crosses between Rhode Island Reds and Barred Plymouth Rocks were used. They received a complete growing ration which is manufactured commercially. The chickens in each experiment were all of the same "breed" and from the same hatch. The males and females were as
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
The Effects of Fasting Upon Plasma Enzyme Levels in Chickens
995
PLASMA ENZYME LEVELS TABLE 1.—The effects of withdrawal of feed upon plasma enzyme levels in chickens
Age
Treatment
Plasma enzyme levels (mean±S.D.)
No. birds
LDH
MDH
GOT
ALD
alkPH
Fasted Control
11 10
1,202 + 192 1,777 + 483
No feed 24 hours 734 + 222 833 ±229
355 + 152 363 + 185
35.2 + 3.5 35.5±5.6
not done
9 wk.
Fasted Control
7 7
989 + 267 1,028 + 205
635 + 117 702 ±144
370+163 340 ±180
37.0 + 2.6 39.7±2.2
not done
13 wk.
Fasted Control
13 14
l,319+448f 847 + 221
1,076 ±304 852 ±288
221± 48 186± 38
18.1+6.2 23.4±5.7
not done
10 wk.
Fasted Control
16 17
885 + 259* 706 + 188
745 + 150 671 + 177
151± 29 156+ 34
18.5 + 4.8 16.2±5.6
72 + 60 119±90
16 wk.
Fasted Control
12 12
258 + 168 267± 99
35.8 + 4.8f 30.0±3.3
not done
10 wk.
Fasted Control
14 14
886 + 279f 549 + 147
710+140+ 403 ± 71
144+ 30* 118+ 31
28.5 + 5.7f 18.8 + 5.7
not done
10 wk.
Fasted Control
14 15
l,154±268t 826 + 228
l,232±325t 851 ±230
140+ 36 154± 45
30.4 + 3.Of 25.5±2.6
not done
10 wk.
Fasted Control
16 17
l,229+389t 706 ±188
973 +285 f 671 ±177
173+ 48 156± 34
20.4 + 6.4 16.2±5.6
59 ±28* 119±90
No feed 48 hours 1,139 ±249* l,848 + 588f 886 ±224 1,089±264
*P<.05. fP<.01. Plasma enzyme levels are expressed as enzyme units per ml. of plasma. One LDH, MDH, or GOT unit is equivalent to the amount of enzyme activity which will cause a decrease of 0.001 in optical density per minute under standard conditions. ALD activity is expressed in dihydroxyacetone units. AlkPH activity is expressed in King-Armstrong units per 100 ml.
nearly equally divided as possible between the fasted and control groups. In four of the experiments, one group of normal birds served as controls for more than one group of fasted birds. Heparinized blood samples were collected by cardiac puncture. The alkPH activity was assayed by a modification of the King-Armstrong technique (Powell and Smith, 1954) on an Auto-Analyzer. It was necessary to dilute the plasma samples one to five for this test. The ALD activity was measured by the method of Friedman and Lapan (1958) on a Klett Summerson photoelectric colorimeter. The LDH, MDH, and GOT assays were conducted on a Beckman DU spectrophotometer. The methods for measuring LDH and MDH levels were previously described (McDaniel and Chute, 1961). The
GOT activity was assayed by the method of Karmen (1955) as described in Sigma Technical Bulletin No. 410 (1958). In the latter test, 0.1 ml. of plasma and 400 units of malic dehydrogenase were used. The data were analyzed by means of the "t" test (Snedecor, 1956). RESULTS The effects of the withdrawal of feed upon the plasma enzyme levels are given in Table 1. The removal of feed for 24 hours did not affect the MDH, GOT, and ALD levels, slightly decreased the alkPH levels, and occasionally increased the LDH levels. The removal of feed for 48 hours, however, caused the LDH, MDH, and ALD levels to increase and the alkPH levels to decrease. The GOT levels were signifi-
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
8 wk.
996
L. S. MCDANIEL AND H. A.
DEMPSEY
TABLE 2.—The effects of withdrawal of water upon plasma enzyme levels in chickens Age
No. Treatment birds
Plasma enyzme levels (mean + S.D.) ALD
alkPH
No water 24 hours 472 + 108* 126 ± 3 4 371± 75 127 + 22
19.0±1.8f 15.2±3.2
not done
LDH
MDH
GOT
Fasted Control
11 11
542 ±157 452+ 86
16 wk.
Fasted Control
16 15
679 + 152 732 + 127
612 + 166f 407± 88
108 + 23 104 ±32
16.6+4.5* 13.6 + 2.4
not done
5wk.
Fasted Control
32 31
691 + 168f 1,165 ±270
954±154f 733 ±150
190+32 192 + 33
18.5±2.8t 16.0±2.6
141 + 101 113 ± 68
10 wk.
Fasted Control
11 11
164±32f 127 + 22
23.9±4.1f 15.2±3.2
not done
6 wk.
Fasted Control
23 22
864±235f 1,165 + 204
870 + 151 941 ±126
181+34* 156 ±29
19.1+3.6 20.1±2.9
203 ±164 223 + 123
5wk.
Fasted Control
33 32
980 + 263 1,034 + 199
901 + 164 904 ±155
191 ± 4 1 184 + 32
21.5±4.7* 17.8 + 5.0
136 +531 201±108
10 wk.
Fasted Control
12 11
169 + 33f 127 ±22
37.9 + 5.3f 15.2±3.2
not done
No water 48 hours 520 + 150 475 + 143* 371± 75 452+ 86
No water 72 hours 457 ±149 446 + 113 452+ 86 371± 75
*P<.05. fP<.01. Enzyme activity units same as used in Table 1.
cantly elevated in only one of the four experiments. The effects of lack of water upon the plasma enzyme levels are presented in Table 2. Withdrawal of water for 24 hours caused an increase in the MDH activity, but the levels of this enzyme returned to normal after 48 or 72 hours. In general, water deprivation caused the LDH levels to decrease and the ALD levels to increase. The alkPH levels were depressed and the GOT levels were elevated when the chickens were fasted for 48 hours. The data in Table 3 indicate that removal of both feed and water for periods up to 48 hours did not affect the LDH, MDH, or GOT levels. The fasting did cause elevated ALD levels and slightly depressed alkPH levels. These results were reversed when chickens were given access to feed and water for 24 hours after they had been deprived of feed and water for
48 hours. Elevated LDH, MDH, and GOT levels and normal ALD and alkPH levels were detected. No statistically significant differences were noted between the plasma enzyme levels of the males and females within the fasted and control groups. Since more than one "breed" was used in these experiments, statistical analysis of the relationship of age to tolerance to fasting was not considered valid although similar results were obtained with all of the different "breeds." DISCUSSION
The standard deviations listed in the tables indicate the considerable individual variation in plasma enzyme levels which exists among chickens. They also indicate an apparent individual variation in the ability of the avian metabolism to withstand fasting. These observations agree with those of Stutts et al. (19S7) on a non-in-
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 28, 2015
10 wk.
997
PLASMA ENZYME LEVELS TABLE 3.—The effects of withdrawal of feed and water upon plasma enzyme levels in chickens
Age
Treatment
No. birds
Plasma enzyme levels (mean±S.D.) LDH
MDH
GOT
ALB
alkPH
Fasted Control
11 12
No feed
22.3 + 3.3 21.2±3.3
not done
7wk.
Fasted Control
17 17
1,019 ±233 1,088 ±276
743 + 139 776 + 139
183 ± 34 191 ± 47
20.1±2.4 19.2±3.2
102± 47 135± 86
7wk.
Fasted Control
21 21
793 ±187 677 ±188
662 ±114 626 ±173
139 + 50 146 + 29
20.1±9.6f 13.2±4.9
96 ± 38 141 ± 83
7wk.
Fasted Control
17 17
18.9±3.2 19.2±3.2
105 ±123 135± 86
7wk.
Fasted Control
21 21
855 ±266 677 ±188
716 + 169 626 + 173
125 ± 39 146 ± 29
18.6±7.1f 13.2±4.9
80 + 22* 141 ± 83
7 wk.
Fasted Control
21 21
1,104 + 285 1,108 ±238
1,003 + 199 893 + 164
278 + 149 223+ 86
23.2 + 8.2f 13.9+4.4
97 ± 70 185 ±189
10 wk.
Fasted Control
10 wk.
Fasted Control
17 17
l,193±414f 752 ±221
877 + 257f 622 + 212
151 + 33* 125 + 30
24.5 + 4.6 20.7+4.6
93+ 37 81 ± 31
10 wk.
Fasted Control
28 30
1,103 + 249 1,014 + 222
801 + 170 830+139
136+14* 124+ 27
15.7 + 2.5 17.4 + 6.2
79 ± 31 85 ± 42
No feed
Access to feed and watei• for 24 hours after fasting 48 hours 837±217f 162+ 35 13.2 + 2.6 20 l,176 + 452t 20 662 + 113 557 + 118 152+ 32 11.6 + 1.7
90+ 38 95+ 55
*P<.05. fP<.01. Enzyme activity units same as used in Table 1.
bred strain of White Leghorns. These factors probably account for the apparent discrepancies in the data presented in this paper. Each enzyme studied appeared to follow an individual trend during fasting. Distinctive trends for enzymes have also been observed in rat livers during fasting (Soberon and Sanchez, 1961). It is interesting that the removal of both feed and water appeared to affect the plasma enzyme levels less than the removal of either feed or water alone. This was especially true in the case of the LDH, MDH, and GOT levels. Twenty-four hours appeared to be a sufficient length of time to allow the alkPH and ALD activities to return to the normal levels after the birds had fasted. On the
other hand, the sudden intake of nutrients appeared to disturb the LDH, MDH, and GOT levels more than the withdrawal of feed and water. If blood samples were collected after the birds were given access to feed and water for a longer period of time, the levels of these enzymes probably would have been normal. Since birds possess a rapid metabolism, it might be assumed that fasting would markedly affect blood enzyme levels. The results of these investigations disagree with this assumption. Even in the experiments where statistically significant differences in the average plasma enzyme levels were found, the ranges of the enzyme levels of the fasted birds and the controls overlaped. In physiologic or genetic studies, the effects
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L. S. MCDANIEL AND H. A. DEMPSEY
of fasting could be important factors. In pathologic studies, however, the enzyme levels of the affected chickens would have to be beyond the range of the enzyme levels of the normal birds to be considered significant. Therefore, anorexia probably would not appreciably affect the results of experiments on plasma enzyme levels in chickens during a pathologic condition.
Studies on the effects of the removal of feed and/or water upon plasma enzyme levels in chickens were conducted. During the three fasting conditions, the aldolase (ALD) levels were usually increased and the alkaline phosphatase (alkPH) levels were usually decreased. No statistically significant differences were noted in the alkPH levels until the birds had been fasted 48 hours. Lack of water affected the ALD levels more than the other two fasting conditions. The withholding of both feed and water for 48 hours did not affect the lactic dehydrogenase (LDH), malic dehydrogenase (MDH), and glutamic oxalacetic transaminase (GOT) levels. The LDH levels usually increased when feed was withheld and usually decreased when water was withheld. The MDH levels were not affected when feed was withheld for 24 hours, but were elevated when feed was withheld for 48 hours. Removal of water for 24 hours caused the MDH levels to increase, but the levels tended to return to normal after water had been removed for 48 hours or more. The GOT levels were elevated when water was withheld for 48 hours, but they were usually not affected when feed was withheld. When chickens were given access to feed
REFERENCES Friedman, M. M., and B. Lapan, 1958. Serum aldolase in the neonatal period: including a colorimetric determination of aldolase by standardization with dihydroxyacetone. J. Lab. Clin. Med. S i : 745-752. Gilfillan, R. F., D. F. Holtman and R. T. Ross, 1956. Influence of Salmonella pullorum infection on various liver tricarboxylic acid enzymes and citrate levels in the chick. J. Bact. 72: 624-627. Hill, C. H., H. W. Garren, J. W. Kelly and M. K. Warren, 1960. Studies on blood and liver citric acid levels in chicks. Poultry Sci. 39: 117-119. Karmen, A., 1955. A note on the spectrophotometric assay of glutamic oxalacetic transaminase in human blood serum. J. Clin. Invest. 34: 131-133. McDaniel, L. S., and H. L. Chute, 1961. Enzyme activity levels in chicken plasma. Am. J. Vet. Res. 22: 99-103. Powell, M. E. A., and M. J. H. Smith, 1954. The determination of serum acid and alkaline phosphatase activity with 4-aminoantipyrine (A.A.P.). J. Clin. Path. 7: 245-248. Sigma Technical Bulletin No. 410, 1958. Determination of glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase at 340mn in serum or other fluids. Sigma Chemical Co., St. Louis, Mo. Snedecor, G. W., 1956. Statistical Methods. 5th Ed. Iowa State College Press., Ames, la. Soberon, G., and E. Sanchez, 1961. Changes in effective enzyme concentration in the growing rat liver. I. Effects of fasting followed by repletion. J. Biol. Chem. 236: 1602-1606. Stutts, E. C , W. E. Briles and H. O. Kunkel, 1957. Plasma alkaline phosphatase activity in mature inbred chickens. Poultry Sci. 36: 269276. Szepsenwol, J., and J. V. Michalski, 1951. Glycogenolysis in the liver and glycogen body of the chicken after death. Am. J. Physiol. 165: 624-627.
AUGUST 9-15, 1963. SIXTH INTERNATIONAL CONGRESS ON NUTRITION, EDINBURGH, SCOTLAND
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SUMMARY
and water for 24 hours after they had been denied both feed and water for 48 hours, elevated LDH, MDH, and GOT levels and normal ALD and alkPH levels were detected.