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Plasma Proteins of Chickens as Influenced by Time of Laying, Ovulation, Number of Blood Samples Taken and Plasma Volume1
Plasma Proteins of Chickens as Influenced by Time of Laying, Ovulation, Number of Blood Samples Taken and Plasma Volume 1 PAUL D. STUKKIE
Laboratory of Avian Physiology, Rutgers University, New Brunswick, N. J. (Received for publication August 22, 1950)
I
N REVIEWING the literature upon plasma or serum proteins of chickens one is impressed by the considerable variation in values reported by the different investigators. Many of the studies have been based upon small numbers and give no details concerning the age, sex, state of reproduction, or other conditions. A number of workers have reported upon total proteins of the blood of chickens of various ages and conditions, but fewer individuals have studied the individual components of the proteins. Howe (1925), in reviewing some of the earlier work, reported that most of the values were too high because of the unreliability of the methods used. He gives a value for total serum proteins of 3.6 grams percent for two adult chickens, of unknown sex. The A/G ratio was 1.02. Other workers, including Dyer and Roe (1934), Chorine (1938), and Defalco (1942) reported A/G ratio (albumin to globulin) of 0.7 to 0.8 with figures for total proteins ranging from 3.3 to 4.8. The exact age and condition of the birds studied by some of these workers were not given. Sturkie (1947),
using the less reliable specific gravity technique, reported total plasma proteins of laying birds of 4.38 grams percent. Rochlina (1934a) calculated the total proteins of laying and non-laying hens based upon the refractive index of the plasma. She reported an average value of 5.54 for total proteins for ten birds, which included laying and non-laying birds. She stated that the values were about 25 to 60 percent lower on the day that the birds laid. She (1934b) showed that when blood samples were taken in the forenoon, noon and afternoon, plasma proteins decreased with succeeding samples. In view of the reports of Rochlina, it was considered desirable to study in more detail the effects of ovulation, laying, and other factors on total proteins and on the individual components, albumin and globulin. Another objective of this experiment was to determine whether the apparent diurnal variation in plasma proteins reported by Rochlina is due to hemodilution and the amount of blood taken at each sample and/or changes in plasma volume. EXPERIMENTAL PROCEDURE
1
Paper of the Journal Series, New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick, New Jersey, Department of Poultry Husbandry.
Details as to age, state of reproduction and other conditions of birds upon which plasma proteins, blood volume, and cell volume were determined are treated in
240
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WITH THE TECHNICAL ASSISTANCE OE H. J. NEWMAN
241
FACTORS INFLUENCING PLASMA PROTEINS RESULTS
Egg production and plasma proteins.—• Total plasma proteins, albumins, and globulins were determined on 23 laying White Leghorn hens in June, 1949 (age 15 to 17 months). Blood samples were taken at about 1 to 3 p.m. on the day that the birds (1) laid only, (2) ovulated only, (3) laid and ovulated, and (4) neither laid nor ovulated. All birds were laying at 50 to 70 percent and none failed to lay for more than 5 days. The results are shown in Table 1. The values are not paired for
TABLE 1.—Plasma proteins, gm./lOO ml., of laying White Leghorn hens Group 1
2
3
4
Laid only on day sample was taken
Ovulated only on day sample was taken
Laid and ovulated on day sample was taken
Neither laid nor ovulated on day sample was taken
Glob. T. P.
Glob. T. P.
Alb.
Alb.
Glob.
19
19
19
2.48
4.93
2.25
2.68
0.12
0.165 0.083 0.13
Glob. T. P.
T. P.
Alb.
23
23
23
20
20
20
21
21
21
Mean
4.64
2.15
2.48
4.88
2.36
2.52
4.75
2.27
Standard error
0.143 0.07
0.13
0.144 0.09
0.12
0;i35 0.08
No. of birds in category
A/G
0.86
Alb.
0.93
plasma and the Klett-Summerson colorimeter were used. Comparison of the micro-Kjeldahl with this method for chicken blood showed agreement within .06 to .08 percent. Plasma volume was determined by using the dye (T-1824) method of Benditt et al., modified as follows: 1 ml. of dye (2.5 mgs.) was injected intravenously and, after a mixing time of 3 minutes, blood samples were taken from the heart. Pino and Weiss (1950) have shown that even 2 minutes is adequate for complete mixing. Concentration of the dye in the plasma and standards was determined with the Klett-Summerson colorimeter. Wintrobe tubes were used in determining hematocrit.
0.91
0.84
each bird, as the same birds were not used in all the groups. The " t " values obtained indicate that none of the differences between these groups, with respect to plasma albumin and globulin were significant. However, the values for the birds, which neither laid nor ovulated were the highest for total protein. The plasma globulins in all groups were higher than the albumins, with A/G ratios ranging from 0.84 to 0.93. In the previous experiment all blood samples were taken in the afternoon, and no attempt was made to study the plasma proteins at the approximate hour of ovulation or at the time, for example, when the rate of deposition of albumin is greatest. Thus, in a later experiment, plasma proteins were determined on laying birds (12
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sections of this paper where they apply. Total plasma proteins and albumin were determined by the biuret method of Gornall el al. (1949). Globulin was determined indirectly. These workers compared this method with the microKjeldahl method on rat serum and found it to agree within 0.1 gram percent of the latter method. They used 0.5 ml. serum or plasma, from which they determined directly the total protein and albumin, colorimetrically, with the Evelyn colorimeter. In the present study 1 ml. of
242
PAUL D. STURKIE AND H. J. NEWMAN
regardless of whether the determinations were made in the forenoon (first sample), noon, or afternoon. It is interesting to note, however, that there was a progressive decrease in plasma proteins with the number of samples taken. This will be discussed later. In the experiments described above, all birds were laying from 50 to 70 percent and none failed to lay for longer than 5 days. In a third experiment, plasma pro.
TABLE 2.—Plasma proteins, gm./lOO ml., ^n relation to time of ovulation and laying Second sample
First sample
Third sample
T. P.
Alb.
Glob.
T. P.
Alb.
Glob.
T. P.
Alb.
Glob.
11
10
10
11
11
11
11
11
11
4.71
2.10
2.64
4.65
2.11
2.55
4.38
2.01
2.32
4
4
11
11
11
11
11
11
4.56
2.08
2.44
4.63
2.10
2.53
4.22
1.82
2.39
12
6
6
12
6
6
12
6
4.55
2.09
2.58
4.54
4.12
2.11
2.27
—
—
No. birds 1* Mean No. birds
6
2 Mean No. birds
6
3 Mean No. birds
2.54
5
5
4 4.23
Mean
2.17
—
—
—
—
—
3.90
* 1 Laid and ovulated; 2 Laid only; 3 Ovulated only; 4 Neither laid nor ovulated.
reached the uterus (3 to 4 p.m.). The results are shown in Table 2. The numbers involved for each of the categories is "in some cases small, and the data were not analyzed statistically, but here again, as was shown in Table 1, there appears to be no difference in plasma proteins that can be related to time of ovulation, or laying, TABLE 3.—Plasma proteins in gm./lOO ml. of IS non-laying While Leghorn hens (16 to 18 months of age) Total protein
Albumin
Globulin
Mean
5.34
2.00
3.34
a mean
0.20
0.09
0.16
A/G
0.60
teins were determined on a group of birds which had not laid for at least 1 month (and in most cases 2 or more months) prior to time blood samples were taken. These birds were White Leghorns, 16 to 18 months of age, and the protein determinations were made in late August of 1949. The results are shown in Table 3. The mean total protein of 5.34 is higher than the values shown in Tables 1 and 2 for laying birds and is significantly so when compared to the lower figures shown in Table 1, but this figure is not higher than that for two other groups of laying birds, whose total protein values averaged 5.18 (Table 6) and 5.32 (Table 7). Thus, these data do not show a con-
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to 15 months of age) divided into the four, groups, as before, with respect to day laid, day ovulated, day laid and ovulated and day neither laid nor ovulated, and also at three different periods of the day. Blood samples were taken at (1) about the time of ovulation for most birds (9 a.m.), (2) at about the time albumin is being, or has been secreted, on the egg in the magnum (11 a.m. to 12 noon), and (3) at approximately the time the egg has
FACTORS INFLUENCING PLASMA PROTEINS
sistent difference in the plasma proteins of laying and non-laying birds. It is noteworthy that the A/G ratio for the nonlaying birds (Table 3) of 0.60 (increased globulins) is considerably lower than the ratios for the other groups of birds. The ratios for all of the other groups of females ranged from 0.83 to 0.93. Further studies on non-laying birds are necessary before
Total protein
Albumin
Globulin
Mean
4.00
1.66
2.33
a mean
0.11
0.06
0.041
A/G
0.71
definite conclusions regarding A/G ratio can be drawn. Plasma proteins of males.—Plasma proteins were determined on 14 adult White Leghorn males (age 18 to 22 months) in July. The results are shown in Table 4. The mean for total protein is 4.00 grams percent, and for albumin and globulin, 1.66 and 2.33 respectively. These values are significantly lower than any of the values for hens. Diurnal variations in plasma proteins.— This experiment was designed to check
the report of Rochlina that plasma proteins are highest in the forenoon and decrease in the afternoon. In this study blood samples were taken from 15 laying White Leghorn hens three times on a given day at the hours already indicated. The data are shown in Table 5. It is observed that there was a progressive decrease in proteins of the second and third samples and that the differences between samples 1 and 3 and 2 and 3 but not 1 and 2 for total protein are significant. The greater part of the decrease in total proteins is due to a decrease in globulin. The actual decrease in total protein between samples 1 and 3 and 2 and 3 are 8.07 and 5.05 percent respectively, and for globulin 11.7 and 10.2 percent respectively. These data confirm the data of Rochlina and might be interpreted to mean a true diurnal variation in plasma proteins. Another experiment was run in which the first blood samples were taken in the afternoon (3 to 4 p.m.) and the second sample was taken the following'morning on most of the same birds. In both experiments a total of 5 cc. of blood was taken from the heart for each analysis. The data are summarized in Table 6. It is observed that the second sample taken at 9 a.m. shows a significant de-
TABLE 5.—Effect of time of blood sampling upon total plasma proteins, albumin, and globulin, in gm./lOO ml. First samples taken at about 9 a.m.; second samples 11 to 12 a.m.; and third samples 3 to 4 p.m. Samples are paired and the difference treated as a mean
Actual means A/G Md a md "t" % change Direction of + change — (number) 0
T. P.
Alb.
4.71
2.14
2 and 3 (3-2)
1 and 3 (3-D
1 and 2 (2-1)
Sample No.
Glob.
T. P.
Alb.
2.57
4.66
2.16
.83 0.38 0.02 0.60 0.08 0.05 0.046 0.049 0.133 0.041 0.076 2.00 0.65 0.43 1.22 2.85 - 1 . 0 7 + .03 - 1 . 5 6 - 8 . 0 7 - 3 . 7 4 7 4 1 4. 7 14 11 8 6 8 — 2 3 — —
.86
Glob.
T.P.
Alb.
2.51
4.33
2.06
Glob. 2.27 90
0.24 0.30 0.33 0.10 0.099 0.136 0.053 0.105 2.42 1.89 2.42 2.85 - 11.70 - 5 . 0 5 - 4 . 6 3 --10.20 2 1 1 1 12 12 13 14 2 1 1 —
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TABLE 4.—Plasma proteins, in gm./lOO ml., of 14 White Leghorn males (18 to 22 months of age)
243
244
PAUL D. STURKIE AND H. J. NEWMAN
TABLE 6.—Summary of results of first and second samples of blood for plasma proteins, in gm./lOO ml., for laying White Leghorn hens. First sample taken 3 to 4 p.m. and second sample the following morning. Values paired Sample 1 2
Actual Means
+ 0
Albumin
5.18 4.85 .33 0.0816 4.1 6.4 10 2 0
2.50. 2.33 .17 0.0451 5.2 6.8 10 2 0
Globulin 2.67 2.51 .16 0.0476 3.3 6.0 10 1 1
A/G 0.93 0.93
crease in albumin and globulin over the first sample taken in the afternoon. The percent decrease for total proteins, 6.2, is of about the same order (slightly higher) as shown by the data in Table 5, where the first sample was taken in forenoon. The percentage decrease in albumins and globulins was about the same. These re^ suits are just the reverse of those of the preceding experiment and show that succeeding blood samples taken from the same bird per day exhibit a significant decrease in plasma proteins regardless of time of day when samples are taken. This was further demonstrated in two other experiments where the first blood samples were taken in the afternoon and where also the second samples were significantly lower in plasma proteins than the first (Tables 7 and 8). In three of the experiments (Tables 5, 6, and 7), 5 to 7 | cc. of TABLE 7.-
TABLE 8.—Differences in total plasma proteins, in gm./lOO ml., of first and second samples taken in afternoon (first) and following morning (second) on i' birds. Values paired
Sample
Total protein
Md (1-2)
a md
"t"
1 2
4.62 4.21
0.449
0.1903
2.36
Direction of change (number)
+
-
0
8
1
0
4.71, is lower than that for the first samples taken in the afternoon, 5.18 and 5.32. The data shown in Table 7 include determinations made on some of the birds, but with additional ones, and those shown in Table 8 pertain to a new group of birds. The means for those groups for total proteins on the first samples were 5.32 and 4.62 respectively. These data, though suggestive, do not prove the existence of a diurnal variation in plasma proteins, independent of changes in blood volume. Accordingly, another experiment was planned using
-Differences in plasma proteins, blood volumes and hematocrit of first and second samples taken in afternoon (first) and morning (second) of laying White Leghorns. Values paired
T. P., gm./lOO ml.
First sample Second sample M d (sample 2-1) (7 md "t" % change Direction of + change — (number) 0
Alb., gm./lOO ml.
Glob., gm./lOO ml.
5.32 5.07
2.53 2.40
2.79 2.67
-0.24 0.078 3.08 -4.70 1 15 0
-0.13 0.042 3.09 -5.14 3 13 0
-0.12 0.059 2.03 -4.30 4 • 12 0
* Adjusted for amount of blood taken onfirstsample.
A/G
Plasma volume ml.
Plasma volume adjust.,* ml.
0.90 0.89
93.56 97.80
88.93 97.80
4.24 1.87 2.26 4.53 5 11 0
8.87 1.87 4.74 9.97 0 16 0
Circulating total plasma proteins, gms. 5.00 4.97 -0.037 0.086 0.446 -0.060 9 7 0
Circulating total plasma proteins adjusted
Cell volume hematocrit, gm./lOO ml.
Cell volume total circulating
28.5 26.4
38.19 35.69
4.64 4.97 0.33 0.079 4.18 7.08 . 2 14 0
-
2.10 0.74 2.80 7.37 12 4 0
-
2.52 1.49 1.69 6.60
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Md (j md 't" % change Direction of change (number)
Total protein
blood were taken on the first sample. In another (Table 8) only 2 cc. were taken. Thus, among the four experiments the first blood sample was taken in the afternoon for three and in the morning for one (Table 5). The data shown in Tables 5 and 6 were on the same birds, but with a period of 2 months intervening between sampling. The mean total proteins for the first sample taken in the morning (Table 5),
FACTORS INFLUENCING PLASMA PROTEINS
highly significant. The hematocrit of the second sample, 26.4, showed a significant decrease of 7.4 percent over the first sample. The total circulating plasma proteins (unadjusted) for the first blood sample in the table are calculated on the basis of proteins per unadjusted total plasma volume (93.5). This gives a figure of 5.00 gms. for the first sample and 4.97 gms. for the second, and this difference is not TABLE 9.—Total plasma proteins, in gm./lOO ml., of laying White Leghorn females at 9 a.m. and 3 to 4 p.m. First sample taken in a.m. and one week later next sample taken in p.m. Values paired Sample
No. Total of proteins birds (means)
1
20
4.61
2
20
4.70
M , /i_i-i '
0.09
"
m
d
0.12
"t"
0.72
significant. Since the plasma volume following the first sampling of blood is decreased by the amount of plasma contained in the first sample of whole blood, the difference in these figures (5.00 and 4.97) does not represent the actual difference between the total circulating plasma proteins of the first and second samples. The adjusted total circulating plasma proteins, which takes into account the changed plasma volume, following the first sample, represents the actual change between the first and second samples. Thus, this adjusted figure for the first sample is 4.64 and for the second, 4.97 gms. The difference of -f 0.33 gms. (7 percent) is highly significant statistically. These data show that with successive samples, plasma proteins in gm./lOO ml. and hematocrit are significantly decreased and that plasma volume is increased. The decrease in plasma proteins is about 5 percent and in hematocrit, 7 percent, and the increase in plasma volume is about 10
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another group of laying birds with blood samples taken in the morning (9 a.m.), and one week later another first sample taken in the afternoon (3 to 4 p.m.). The data are shown in Table 9. It is observed that the difference between total plasma proteins in the morning and afternoon (0.087 gram percent) is not significant. These data piove conclusively that between the daylight hours of 9 a.m. and 4 p.m. there is no true variation in plasma proteins per bird. Plasma proteins influenced by changes in blood volume.—The fact that successive blood samples from the same bird per day showed decreased plasma proteins suggested the possibility that sampling may have produced hemodilution and increased plasma volume. Thus, an experiment was planned in which plasma proteins, hematocrit and blood volume were determined on two blood samples per bird, the first being taken in the afternoon (3 to 4 p.m.) and the second one the following morning at 9 a.m. A total of 7§ cc. of blood were taken per bird per sample. Two cc. were taken first from the wing vein, then 1 cc. of T-1824 dye (2.5 mgs.) was injected intravenously. After a mixing time of 3 minutes, 5 to 5 | cc. were taken from the heart. The data are summarized in Table 7. The total protein, albumin and globulin of the second sample as in other experiments, were significantly decreased, approximately 5 percent. The actual plasma volume of the first sample was 93.5 cc. and that of the second sample was 97.8 cc. The increase is greater than indicated, however, because after the first sampling the volume remaining was decreased by the amount of plasma contained in 6J cc. of whole blood. Thus, the corrected plasma volume of 88.93 is subtracted from 97.8 to give the actual increase in volume, which amounts to approximately 10 percent. This difference is
245
246
PAUL D. STURKIE AND H. J. NEWMAN
DISCUSSION The results of this study show that time of ovulation, time of laying, and position of egg in oviduct at the time of determinations had no significant effect upon plasma albumin or globulin. While the plasma proteins of persistent nonlaying birds appeared higher than some of the laying birds in this study, the figure was no higher than that reported for some groups which were laying. These results do not support those of Rochlina. The A/G ratio of the laying birds studied ranged from 0.83 to 0.93, while the A/G ratio of the one group of non-laying birds was 0.60. The plasma protein of the males was significantly lower than those of the females, and the A/G ratio was 0.70. The data show that time of sampling, frequency of sampling, time of ovulation, or oviposition, and changes in blood volume did not appreciably affect the A/G ratios of the laying birds. Levin and Leathern (1942) suggested that in the rat the serum albumin level is maintained under the influence of the adrenal cortical hormones, while the serum globulin level- is associated with
thyroid activity. They showed that administration of stilbesterol to intact rats increased plasma albumin but not globulin, presumably via the adrenal cortex. Thus, if this mechanism of control applies in the chicken one might expect that plasma albumin in the chicken would be lower (lower A/G ratio) in non-laying females and in males, where presumably circulating estrogens are low or absent. In the pigeon, which has a much higher A/G ratio than the chicken (greater than 1) administration of estrogens to intact females increases total proteins and mainly through an increase in albumin (Mandel et ah, 1947). Sturkie (unpublished), who administered estrogens to laying females, found that though total proteins were increased considerably, there was no significant change in the A/G ratio. Further studies on the effects of hormones on the components of the plasma proteins of the chicken are necess'ary before conclusions can be made concerning the mechanism by which the normal levels of albumin and globulin are maintained. The results of the effects of time and frequency of blood sampling on changes in blood volume are noteworthy. It is recalled from Table 1 that when three successive samples are taken from the same bird per day, the second sample, taken 2 to 3 hours after the first, showed a slight but insignificant decrease in protein, but the third sample, taken 3 to 4 hours later, showed a significant decrease in proteins over the first and second samples. This suggests that the act of taking the sample (in this case heart puncture) sets in motion a shift of water from the tissues to the blood, resulting in increased plasma volume and decreased cellular volume, but that the effect of the shift is not appreciable until after 5 or 6 hours. It is noted that this effect still exists after
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percent. Thus, the decrease in protein in grams percent is due mainly to an increase in volume. In terms of total circulating plasma proteins, however, there is an actual increase in the second sample. The total circulating cell volume, which is calculated from corrected plasma volume and hematocrit, still shows a decrease of about the same order as the hematocrit for the second sample. These data are interpreted to mean that following the hemodilution and increased plasma volume, occasioned by frequent sampling, there occurred an actual increase in the synthesis of plasma proteins (total circulating plasma proteins) but not in the formation of corpuscles.
FACTORS INFLUENCING PLASMA PROTEINS
It is important to note, however, that stress induced in chickens by hypothermia increases plasma proteins and causes hemoconcentration and presumably decreases plasma volume (Sturkie, 1947), although volume was not determined. It is known, moreover, that different types of stress do not always elicit the same response in some species. Further work is necessary to determine the mechanism by which plasma volume changes are controlled, since such changes may influence not only determinations of plasma proteins but other constituents as well. SUMMARY
The results of this study show that time of ovulation, time of laying, and position of the egg in the oviduct at the time of determinations had no significant effect upon plasma albumin and globulin. The A/G ratios of the laying birds studied
ranged from 0.83 to 0.93 an.d that of the one group of non-laying birds was 0.60. The plasma proteins of the males were significantly lower than those of the females and the A/G ratio was 0.70. Time of sampling, frequency of sampling, time of ovulation or oviposition and changes in blood volume did not appreciably affect the A/G ratios of the laying birds. Between the hours of 9 a.m. and 4 p.m. there is no true variation in plasma proteins of laying birds. When successive samples on the same bird are taken, the second or third samples always show a decrease in plasma proteins over the first sample, regardless of the time of day when the samples are taken. This decrease in plasma proteins was shown to be due to hemodilution and an increase in plasma volume. Also, there was a decrease in hematocrit. Actually, when the plasma proteins are calculated on the basis of adjusted total circulating proteins it is found that there is a slight increase in the synthesis of plasma proteins but not in the formation of the corpuscles of the blood. REFERENCES Benditt, E. P., R. L. Straube and E. M. Humphreys, 1946. The determination of total circulating serum proteins and erythrocyte volumes in normal and protein-depleted rats. Proc. Soc. Exper. Biol. Med. 62: 189-192. Chorine, V., 1938. Proteins of chicken serum. Comp. Rend. Soc. Biol. 127: 170-173. Defalco, R. J., 1942. A seriological study of some avian relationships. Biol. Bull. 83: 205-218. Dyer, H. M., and J. H. Roe, 1934. The chemistry of the blood of normal chickens. J. Nutrition, 7: 623-626. Gornall, A. G., C. J. Bardawill and M. D. David, 1949. Determination of serum proteins by means of the biuret reaction. J. Biol. Chem. 177: 751— 766. Howe, P. E., 1925. The function of the plasma proteins. Physiol. Reviews 5: 439-476. ' Levin, Louis and J. H. Leathern, 1942. The relation of the pituitary, thyroid and adrenal glands to the maintenance of normal serum albumin and globulin levels. Am. J. Physiol. 136: 306-313.
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16 to 18 hours have elapsed between the taking of the first and second samples. It is recalled that the amount of blood taken on the first sample is not the sole factor which initiates the shift, because even when only 2 cc. of blood are taken, the change occurs, though the magnitude of change is not so great as when 5 to 7 cc. are taken. It is possible that the stress induced by cardiac puncture or handling of the bird in taking the blood sample causes the adrenal to release cortical hormones which increase the serum sodium with a consequent increase in plasma volume and hemodilution. It is known that desoxycorticosterone acetate (DCA) produces this effect in certain mammals, and Selye (1942) found that DC A when injected into chicks produced hemodilution and tissue edema. He did not determine blood volume, but he thought that the decrease in hemoglobin was secondary to an increase in blood volume.
247
248
NEWS AND NOTES
Mandel, P., J. Clavert and L. Mandel, 1947. Effects de ia folliculine sur les diverses fractions proteiques du plasma chez le pigeon. Comp. Rend. Soc. Biol. 141:678-680. Pino, J. A. and H. S. Weiss, 1950. A technique for the measurement of blood volume. Poultry Sci. 29: 776. Rochlina, M., 1934a. Les proteines du sang et la ponte des poules. Bull. Soc. de Chemie Biologique 16: 1645-1651.
Rochlina, M., 1934b. Le taux des differents el&nents dans le sang des poules en liaison avec la ponte. Bull. Soc. de Chemie Biologique, 16: 1652-1662. Selye, H., 1942, Production of nephrosclerosis by overdosage with desoxycorticosterone acetate. Canadian Med. Assoc. J. 47: 515-519. Sturkie, P. D., 1947. Effects of hypothermia upon the specific gravity and proteins of the blood of chickens. Am. J. Physiol. 148: 610-613. Sturkie, P. D., 1950. Unpublished.
{Continued from page 230)
E. L. TerBush (B.S.A.—Cornell University) has been appointed Graduate Research Assistant in poultry genetics. H. F. Butlers (B.S.A.—University of Maine) is serving as Graduate Research Assistant in poultry nutrition. C. H. Moore (B.S.A.—Alabama Polytechnic Institute; M.S.—Kansas State College) was transferred from Graduate Research Assistant to Cooperative Agent with the Bureau of Animal Industry. He will assist Dr. Warren in the activities of the Regional Poultry Breeding Project. WASHINGTON NOTES
M. S. Mitchell has been appointed Teaching Assistant in the Department of Poultry Husbandry. He is a graduate of the University of Manitoba, Winnipeg, Canada and will proceed with degree work for the M.S., specializing in poultry genetics. T. Myint (Ph.D.—-Utah State College) has been appointed Research Associate in the Department of Poultry Husbandry, working on the project dealing with the development of microbiological techniques
in the analyses of amino acids and proteins. Dr. Myint is a native of Rangoon, Burma.
E. Sauter (B.S.—Washington State College) has been appointed Research Assistant in poultry products. He will investigate relationships between egg quality and culinary use of the egg, in cooperation with the Department of Foods and Nutrition in the College of Home Economics. OHIO NOTES
Appointments to graduate research assistantships in Poultry Science on projects of the Ohio Agricultural Experiment Station during the academic year 1950-51 are: P. L. Clayton (M.S., 1950, Ohio); L. S. Mercia (B.S., 1950, Vermont) and A. W. Jasper (B.S., 1950, Vermont) in economics and marketing; J. W. Marguand (B.S., 1948, Ohio) in turkey production; Charles McFarland (M.S., 1950, Ohio); D. W. Puffenberger (B.S., 1949, West Virginia) and C. R. Wettling (M.S., 1950, Ohio) in processing preservation of poultry products.
{Continued on page 266)
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News and Notes
Laboratory of Avian Physiology, Rutgers University, New Brunswick, N. J. (Received for publication August 22, 1950)
I
N REVIEWING the literature upon plasma or serum proteins of chickens one is impressed by the considerable variation in values reported by the different investigators. Many of the studies have been based upon small numbers and give no details concerning the age, sex, state of reproduction, or other conditions. A number of workers have reported upon total proteins of the blood of chickens of various ages and conditions, but fewer individuals have studied the individual components of the proteins. Howe (1925), in reviewing some of the earlier work, reported that most of the values were too high because of the unreliability of the methods used. He gives a value for total serum proteins of 3.6 grams percent for two adult chickens, of unknown sex. The A/G ratio was 1.02. Other workers, including Dyer and Roe (1934), Chorine (1938), and Defalco (1942) reported A/G ratio (albumin to globulin) of 0.7 to 0.8 with figures for total proteins ranging from 3.3 to 4.8. The exact age and condition of the birds studied by some of these workers were not given. Sturkie (1947),
using the less reliable specific gravity technique, reported total plasma proteins of laying birds of 4.38 grams percent. Rochlina (1934a) calculated the total proteins of laying and non-laying hens based upon the refractive index of the plasma. She reported an average value of 5.54 for total proteins for ten birds, which included laying and non-laying birds. She stated that the values were about 25 to 60 percent lower on the day that the birds laid. She (1934b) showed that when blood samples were taken in the forenoon, noon and afternoon, plasma proteins decreased with succeeding samples. In view of the reports of Rochlina, it was considered desirable to study in more detail the effects of ovulation, laying, and other factors on total proteins and on the individual components, albumin and globulin. Another objective of this experiment was to determine whether the apparent diurnal variation in plasma proteins reported by Rochlina is due to hemodilution and the amount of blood taken at each sample and/or changes in plasma volume. EXPERIMENTAL PROCEDURE
1
Paper of the Journal Series, New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick, New Jersey, Department of Poultry Husbandry.
Details as to age, state of reproduction and other conditions of birds upon which plasma proteins, blood volume, and cell volume were determined are treated in
240
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WITH THE TECHNICAL ASSISTANCE OE H. J. NEWMAN
241
FACTORS INFLUENCING PLASMA PROTEINS RESULTS
Egg production and plasma proteins.—• Total plasma proteins, albumins, and globulins were determined on 23 laying White Leghorn hens in June, 1949 (age 15 to 17 months). Blood samples were taken at about 1 to 3 p.m. on the day that the birds (1) laid only, (2) ovulated only, (3) laid and ovulated, and (4) neither laid nor ovulated. All birds were laying at 50 to 70 percent and none failed to lay for more than 5 days. The results are shown in Table 1. The values are not paired for
TABLE 1.—Plasma proteins, gm./lOO ml., of laying White Leghorn hens Group 1
2
3
4
Laid only on day sample was taken
Ovulated only on day sample was taken
Laid and ovulated on day sample was taken
Neither laid nor ovulated on day sample was taken
Glob. T. P.
Glob. T. P.
Alb.
Alb.
Glob.
19
19
19
2.48
4.93
2.25
2.68
0.12
0.165 0.083 0.13
Glob. T. P.
T. P.
Alb.
23
23
23
20
20
20
21
21
21
Mean
4.64
2.15
2.48
4.88
2.36
2.52
4.75
2.27
Standard error
0.143 0.07
0.13
0.144 0.09
0.12
0;i35 0.08
No. of birds in category
A/G
0.86
Alb.
0.93
plasma and the Klett-Summerson colorimeter were used. Comparison of the micro-Kjeldahl with this method for chicken blood showed agreement within .06 to .08 percent. Plasma volume was determined by using the dye (T-1824) method of Benditt et al., modified as follows: 1 ml. of dye (2.5 mgs.) was injected intravenously and, after a mixing time of 3 minutes, blood samples were taken from the heart. Pino and Weiss (1950) have shown that even 2 minutes is adequate for complete mixing. Concentration of the dye in the plasma and standards was determined with the Klett-Summerson colorimeter. Wintrobe tubes were used in determining hematocrit.
0.91
0.84
each bird, as the same birds were not used in all the groups. The " t " values obtained indicate that none of the differences between these groups, with respect to plasma albumin and globulin were significant. However, the values for the birds, which neither laid nor ovulated were the highest for total protein. The plasma globulins in all groups were higher than the albumins, with A/G ratios ranging from 0.84 to 0.93. In the previous experiment all blood samples were taken in the afternoon, and no attempt was made to study the plasma proteins at the approximate hour of ovulation or at the time, for example, when the rate of deposition of albumin is greatest. Thus, in a later experiment, plasma proteins were determined on laying birds (12
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sections of this paper where they apply. Total plasma proteins and albumin were determined by the biuret method of Gornall el al. (1949). Globulin was determined indirectly. These workers compared this method with the microKjeldahl method on rat serum and found it to agree within 0.1 gram percent of the latter method. They used 0.5 ml. serum or plasma, from which they determined directly the total protein and albumin, colorimetrically, with the Evelyn colorimeter. In the present study 1 ml. of
242
PAUL D. STURKIE AND H. J. NEWMAN
regardless of whether the determinations were made in the forenoon (first sample), noon, or afternoon. It is interesting to note, however, that there was a progressive decrease in plasma proteins with the number of samples taken. This will be discussed later. In the experiments described above, all birds were laying from 50 to 70 percent and none failed to lay for longer than 5 days. In a third experiment, plasma pro.
TABLE 2.—Plasma proteins, gm./lOO ml., ^n relation to time of ovulation and laying Second sample
First sample
Third sample
T. P.
Alb.
Glob.
T. P.
Alb.
Glob.
T. P.
Alb.
Glob.
11
10
10
11
11
11
11
11
11
4.71
2.10
2.64
4.65
2.11
2.55
4.38
2.01
2.32
4
4
11
11
11
11
11
11
4.56
2.08
2.44
4.63
2.10
2.53
4.22
1.82
2.39
12
6
6
12
6
6
12
6
4.55
2.09
2.58
4.54
4.12
2.11
2.27
—
—
No. birds 1* Mean No. birds
6
2 Mean No. birds
6
3 Mean No. birds
2.54
5
5
4 4.23
Mean
2.17
—
—
—
—
—
3.90
* 1 Laid and ovulated; 2 Laid only; 3 Ovulated only; 4 Neither laid nor ovulated.
reached the uterus (3 to 4 p.m.). The results are shown in Table 2. The numbers involved for each of the categories is "in some cases small, and the data were not analyzed statistically, but here again, as was shown in Table 1, there appears to be no difference in plasma proteins that can be related to time of ovulation, or laying, TABLE 3.—Plasma proteins in gm./lOO ml. of IS non-laying While Leghorn hens (16 to 18 months of age) Total protein
Albumin
Globulin
Mean
5.34
2.00
3.34
a mean
0.20
0.09
0.16
A/G
0.60
teins were determined on a group of birds which had not laid for at least 1 month (and in most cases 2 or more months) prior to time blood samples were taken. These birds were White Leghorns, 16 to 18 months of age, and the protein determinations were made in late August of 1949. The results are shown in Table 3. The mean total protein of 5.34 is higher than the values shown in Tables 1 and 2 for laying birds and is significantly so when compared to the lower figures shown in Table 1, but this figure is not higher than that for two other groups of laying birds, whose total protein values averaged 5.18 (Table 6) and 5.32 (Table 7). Thus, these data do not show a con-
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to 15 months of age) divided into the four, groups, as before, with respect to day laid, day ovulated, day laid and ovulated and day neither laid nor ovulated, and also at three different periods of the day. Blood samples were taken at (1) about the time of ovulation for most birds (9 a.m.), (2) at about the time albumin is being, or has been secreted, on the egg in the magnum (11 a.m. to 12 noon), and (3) at approximately the time the egg has
FACTORS INFLUENCING PLASMA PROTEINS
sistent difference in the plasma proteins of laying and non-laying birds. It is noteworthy that the A/G ratio for the nonlaying birds (Table 3) of 0.60 (increased globulins) is considerably lower than the ratios for the other groups of birds. The ratios for all of the other groups of females ranged from 0.83 to 0.93. Further studies on non-laying birds are necessary before
Total protein
Albumin
Globulin
Mean
4.00
1.66
2.33
a mean
0.11
0.06
0.041
A/G
0.71
definite conclusions regarding A/G ratio can be drawn. Plasma proteins of males.—Plasma proteins were determined on 14 adult White Leghorn males (age 18 to 22 months) in July. The results are shown in Table 4. The mean for total protein is 4.00 grams percent, and for albumin and globulin, 1.66 and 2.33 respectively. These values are significantly lower than any of the values for hens. Diurnal variations in plasma proteins.— This experiment was designed to check
the report of Rochlina that plasma proteins are highest in the forenoon and decrease in the afternoon. In this study blood samples were taken from 15 laying White Leghorn hens three times on a given day at the hours already indicated. The data are shown in Table 5. It is observed that there was a progressive decrease in proteins of the second and third samples and that the differences between samples 1 and 3 and 2 and 3 but not 1 and 2 for total protein are significant. The greater part of the decrease in total proteins is due to a decrease in globulin. The actual decrease in total protein between samples 1 and 3 and 2 and 3 are 8.07 and 5.05 percent respectively, and for globulin 11.7 and 10.2 percent respectively. These data confirm the data of Rochlina and might be interpreted to mean a true diurnal variation in plasma proteins. Another experiment was run in which the first blood samples were taken in the afternoon (3 to 4 p.m.) and the second sample was taken the following'morning on most of the same birds. In both experiments a total of 5 cc. of blood was taken from the heart for each analysis. The data are summarized in Table 6. It is observed that the second sample taken at 9 a.m. shows a significant de-
TABLE 5.—Effect of time of blood sampling upon total plasma proteins, albumin, and globulin, in gm./lOO ml. First samples taken at about 9 a.m.; second samples 11 to 12 a.m.; and third samples 3 to 4 p.m. Samples are paired and the difference treated as a mean
Actual means A/G Md a md "t" % change Direction of + change — (number) 0
T. P.
Alb.
4.71
2.14
2 and 3 (3-2)
1 and 3 (3-D
1 and 2 (2-1)
Sample No.
Glob.
T. P.
Alb.
2.57
4.66
2.16
.83 0.38 0.02 0.60 0.08 0.05 0.046 0.049 0.133 0.041 0.076 2.00 0.65 0.43 1.22 2.85 - 1 . 0 7 + .03 - 1 . 5 6 - 8 . 0 7 - 3 . 7 4 7 4 1 4. 7 14 11 8 6 8 — 2 3 — —
.86
Glob.
T.P.
Alb.
2.51
4.33
2.06
Glob. 2.27 90
0.24 0.30 0.33 0.10 0.099 0.136 0.053 0.105 2.42 1.89 2.42 2.85 - 11.70 - 5 . 0 5 - 4 . 6 3 --10.20 2 1 1 1 12 12 13 14 2 1 1 —
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TABLE 4.—Plasma proteins, in gm./lOO ml., of 14 White Leghorn males (18 to 22 months of age)
243
244
PAUL D. STURKIE AND H. J. NEWMAN
TABLE 6.—Summary of results of first and second samples of blood for plasma proteins, in gm./lOO ml., for laying White Leghorn hens. First sample taken 3 to 4 p.m. and second sample the following morning. Values paired Sample 1 2
Actual Means
+ 0
Albumin
5.18 4.85 .33 0.0816 4.1 6.4 10 2 0
2.50. 2.33 .17 0.0451 5.2 6.8 10 2 0
Globulin 2.67 2.51 .16 0.0476 3.3 6.0 10 1 1
A/G 0.93 0.93
crease in albumin and globulin over the first sample taken in the afternoon. The percent decrease for total proteins, 6.2, is of about the same order (slightly higher) as shown by the data in Table 5, where the first sample was taken in forenoon. The percentage decrease in albumins and globulins was about the same. These re^ suits are just the reverse of those of the preceding experiment and show that succeeding blood samples taken from the same bird per day exhibit a significant decrease in plasma proteins regardless of time of day when samples are taken. This was further demonstrated in two other experiments where the first blood samples were taken in the afternoon and where also the second samples were significantly lower in plasma proteins than the first (Tables 7 and 8). In three of the experiments (Tables 5, 6, and 7), 5 to 7 | cc. of TABLE 7.-
TABLE 8.—Differences in total plasma proteins, in gm./lOO ml., of first and second samples taken in afternoon (first) and following morning (second) on i' birds. Values paired
Sample
Total protein
Md (1-2)
a md
"t"
1 2
4.62 4.21
0.449
0.1903
2.36
Direction of change (number)
+
-
0
8
1
0
4.71, is lower than that for the first samples taken in the afternoon, 5.18 and 5.32. The data shown in Table 7 include determinations made on some of the birds, but with additional ones, and those shown in Table 8 pertain to a new group of birds. The means for those groups for total proteins on the first samples were 5.32 and 4.62 respectively. These data, though suggestive, do not prove the existence of a diurnal variation in plasma proteins, independent of changes in blood volume. Accordingly, another experiment was planned using
-Differences in plasma proteins, blood volumes and hematocrit of first and second samples taken in afternoon (first) and morning (second) of laying White Leghorns. Values paired
T. P., gm./lOO ml.
First sample Second sample M d (sample 2-1) (7 md "t" % change Direction of + change — (number) 0
Alb., gm./lOO ml.
Glob., gm./lOO ml.
5.32 5.07
2.53 2.40
2.79 2.67
-0.24 0.078 3.08 -4.70 1 15 0
-0.13 0.042 3.09 -5.14 3 13 0
-0.12 0.059 2.03 -4.30 4 • 12 0
* Adjusted for amount of blood taken onfirstsample.
A/G
Plasma volume ml.
Plasma volume adjust.,* ml.
0.90 0.89
93.56 97.80
88.93 97.80
4.24 1.87 2.26 4.53 5 11 0
8.87 1.87 4.74 9.97 0 16 0
Circulating total plasma proteins, gms. 5.00 4.97 -0.037 0.086 0.446 -0.060 9 7 0
Circulating total plasma proteins adjusted
Cell volume hematocrit, gm./lOO ml.
Cell volume total circulating
28.5 26.4
38.19 35.69
4.64 4.97 0.33 0.079 4.18 7.08 . 2 14 0
-
2.10 0.74 2.80 7.37 12 4 0
-
2.52 1.49 1.69 6.60
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Md (j md 't" % change Direction of change (number)
Total protein
blood were taken on the first sample. In another (Table 8) only 2 cc. were taken. Thus, among the four experiments the first blood sample was taken in the afternoon for three and in the morning for one (Table 5). The data shown in Tables 5 and 6 were on the same birds, but with a period of 2 months intervening between sampling. The mean total proteins for the first sample taken in the morning (Table 5),
FACTORS INFLUENCING PLASMA PROTEINS
highly significant. The hematocrit of the second sample, 26.4, showed a significant decrease of 7.4 percent over the first sample. The total circulating plasma proteins (unadjusted) for the first blood sample in the table are calculated on the basis of proteins per unadjusted total plasma volume (93.5). This gives a figure of 5.00 gms. for the first sample and 4.97 gms. for the second, and this difference is not TABLE 9.—Total plasma proteins, in gm./lOO ml., of laying White Leghorn females at 9 a.m. and 3 to 4 p.m. First sample taken in a.m. and one week later next sample taken in p.m. Values paired Sample
No. Total of proteins birds (means)
1
20
4.61
2
20
4.70
M , /i_i-i '
0.09
"
m
d
0.12
"t"
0.72
significant. Since the plasma volume following the first sampling of blood is decreased by the amount of plasma contained in the first sample of whole blood, the difference in these figures (5.00 and 4.97) does not represent the actual difference between the total circulating plasma proteins of the first and second samples. The adjusted total circulating plasma proteins, which takes into account the changed plasma volume, following the first sample, represents the actual change between the first and second samples. Thus, this adjusted figure for the first sample is 4.64 and for the second, 4.97 gms. The difference of -f 0.33 gms. (7 percent) is highly significant statistically. These data show that with successive samples, plasma proteins in gm./lOO ml. and hematocrit are significantly decreased and that plasma volume is increased. The decrease in plasma proteins is about 5 percent and in hematocrit, 7 percent, and the increase in plasma volume is about 10
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another group of laying birds with blood samples taken in the morning (9 a.m.), and one week later another first sample taken in the afternoon (3 to 4 p.m.). The data are shown in Table 9. It is observed that the difference between total plasma proteins in the morning and afternoon (0.087 gram percent) is not significant. These data piove conclusively that between the daylight hours of 9 a.m. and 4 p.m. there is no true variation in plasma proteins per bird. Plasma proteins influenced by changes in blood volume.—The fact that successive blood samples from the same bird per day showed decreased plasma proteins suggested the possibility that sampling may have produced hemodilution and increased plasma volume. Thus, an experiment was planned in which plasma proteins, hematocrit and blood volume were determined on two blood samples per bird, the first being taken in the afternoon (3 to 4 p.m.) and the second one the following morning at 9 a.m. A total of 7§ cc. of blood were taken per bird per sample. Two cc. were taken first from the wing vein, then 1 cc. of T-1824 dye (2.5 mgs.) was injected intravenously. After a mixing time of 3 minutes, 5 to 5 | cc. were taken from the heart. The data are summarized in Table 7. The total protein, albumin and globulin of the second sample as in other experiments, were significantly decreased, approximately 5 percent. The actual plasma volume of the first sample was 93.5 cc. and that of the second sample was 97.8 cc. The increase is greater than indicated, however, because after the first sampling the volume remaining was decreased by the amount of plasma contained in 6J cc. of whole blood. Thus, the corrected plasma volume of 88.93 is subtracted from 97.8 to give the actual increase in volume, which amounts to approximately 10 percent. This difference is
245
246
PAUL D. STURKIE AND H. J. NEWMAN
DISCUSSION The results of this study show that time of ovulation, time of laying, and position of egg in oviduct at the time of determinations had no significant effect upon plasma albumin or globulin. While the plasma proteins of persistent nonlaying birds appeared higher than some of the laying birds in this study, the figure was no higher than that reported for some groups which were laying. These results do not support those of Rochlina. The A/G ratio of the laying birds studied ranged from 0.83 to 0.93, while the A/G ratio of the one group of non-laying birds was 0.60. The plasma protein of the males was significantly lower than those of the females, and the A/G ratio was 0.70. The data show that time of sampling, frequency of sampling, time of ovulation, or oviposition, and changes in blood volume did not appreciably affect the A/G ratios of the laying birds. Levin and Leathern (1942) suggested that in the rat the serum albumin level is maintained under the influence of the adrenal cortical hormones, while the serum globulin level- is associated with
thyroid activity. They showed that administration of stilbesterol to intact rats increased plasma albumin but not globulin, presumably via the adrenal cortex. Thus, if this mechanism of control applies in the chicken one might expect that plasma albumin in the chicken would be lower (lower A/G ratio) in non-laying females and in males, where presumably circulating estrogens are low or absent. In the pigeon, which has a much higher A/G ratio than the chicken (greater than 1) administration of estrogens to intact females increases total proteins and mainly through an increase in albumin (Mandel et ah, 1947). Sturkie (unpublished), who administered estrogens to laying females, found that though total proteins were increased considerably, there was no significant change in the A/G ratio. Further studies on the effects of hormones on the components of the plasma proteins of the chicken are necess'ary before conclusions can be made concerning the mechanism by which the normal levels of albumin and globulin are maintained. The results of the effects of time and frequency of blood sampling on changes in blood volume are noteworthy. It is recalled from Table 1 that when three successive samples are taken from the same bird per day, the second sample, taken 2 to 3 hours after the first, showed a slight but insignificant decrease in protein, but the third sample, taken 3 to 4 hours later, showed a significant decrease in proteins over the first and second samples. This suggests that the act of taking the sample (in this case heart puncture) sets in motion a shift of water from the tissues to the blood, resulting in increased plasma volume and decreased cellular volume, but that the effect of the shift is not appreciable until after 5 or 6 hours. It is noted that this effect still exists after
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percent. Thus, the decrease in protein in grams percent is due mainly to an increase in volume. In terms of total circulating plasma proteins, however, there is an actual increase in the second sample. The total circulating cell volume, which is calculated from corrected plasma volume and hematocrit, still shows a decrease of about the same order as the hematocrit for the second sample. These data are interpreted to mean that following the hemodilution and increased plasma volume, occasioned by frequent sampling, there occurred an actual increase in the synthesis of plasma proteins (total circulating plasma proteins) but not in the formation of corpuscles.
FACTORS INFLUENCING PLASMA PROTEINS
It is important to note, however, that stress induced in chickens by hypothermia increases plasma proteins and causes hemoconcentration and presumably decreases plasma volume (Sturkie, 1947), although volume was not determined. It is known, moreover, that different types of stress do not always elicit the same response in some species. Further work is necessary to determine the mechanism by which plasma volume changes are controlled, since such changes may influence not only determinations of plasma proteins but other constituents as well. SUMMARY
The results of this study show that time of ovulation, time of laying, and position of the egg in the oviduct at the time of determinations had no significant effect upon plasma albumin and globulin. The A/G ratios of the laying birds studied
ranged from 0.83 to 0.93 an.d that of the one group of non-laying birds was 0.60. The plasma proteins of the males were significantly lower than those of the females and the A/G ratio was 0.70. Time of sampling, frequency of sampling, time of ovulation or oviposition and changes in blood volume did not appreciably affect the A/G ratios of the laying birds. Between the hours of 9 a.m. and 4 p.m. there is no true variation in plasma proteins of laying birds. When successive samples on the same bird are taken, the second or third samples always show a decrease in plasma proteins over the first sample, regardless of the time of day when the samples are taken. This decrease in plasma proteins was shown to be due to hemodilution and an increase in plasma volume. Also, there was a decrease in hematocrit. Actually, when the plasma proteins are calculated on the basis of adjusted total circulating proteins it is found that there is a slight increase in the synthesis of plasma proteins but not in the formation of the corpuscles of the blood. REFERENCES Benditt, E. P., R. L. Straube and E. M. Humphreys, 1946. The determination of total circulating serum proteins and erythrocyte volumes in normal and protein-depleted rats. Proc. Soc. Exper. Biol. Med. 62: 189-192. Chorine, V., 1938. Proteins of chicken serum. Comp. Rend. Soc. Biol. 127: 170-173. Defalco, R. J., 1942. A seriological study of some avian relationships. Biol. Bull. 83: 205-218. Dyer, H. M., and J. H. Roe, 1934. The chemistry of the blood of normal chickens. J. Nutrition, 7: 623-626. Gornall, A. G., C. J. Bardawill and M. D. David, 1949. Determination of serum proteins by means of the biuret reaction. J. Biol. Chem. 177: 751— 766. Howe, P. E., 1925. The function of the plasma proteins. Physiol. Reviews 5: 439-476. ' Levin, Louis and J. H. Leathern, 1942. The relation of the pituitary, thyroid and adrenal glands to the maintenance of normal serum albumin and globulin levels. Am. J. Physiol. 136: 306-313.
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16 to 18 hours have elapsed between the taking of the first and second samples. It is recalled that the amount of blood taken on the first sample is not the sole factor which initiates the shift, because even when only 2 cc. of blood are taken, the change occurs, though the magnitude of change is not so great as when 5 to 7 cc. are taken. It is possible that the stress induced by cardiac puncture or handling of the bird in taking the blood sample causes the adrenal to release cortical hormones which increase the serum sodium with a consequent increase in plasma volume and hemodilution. It is known that desoxycorticosterone acetate (DCA) produces this effect in certain mammals, and Selye (1942) found that DC A when injected into chicks produced hemodilution and tissue edema. He did not determine blood volume, but he thought that the decrease in hemoglobin was secondary to an increase in blood volume.
247
248
NEWS AND NOTES
Mandel, P., J. Clavert and L. Mandel, 1947. Effects de ia folliculine sur les diverses fractions proteiques du plasma chez le pigeon. Comp. Rend. Soc. Biol. 141:678-680. Pino, J. A. and H. S. Weiss, 1950. A technique for the measurement of blood volume. Poultry Sci. 29: 776. Rochlina, M., 1934a. Les proteines du sang et la ponte des poules. Bull. Soc. de Chemie Biologique 16: 1645-1651.
Rochlina, M., 1934b. Le taux des differents el&nents dans le sang des poules en liaison avec la ponte. Bull. Soc. de Chemie Biologique, 16: 1652-1662. Selye, H., 1942, Production of nephrosclerosis by overdosage with desoxycorticosterone acetate. Canadian Med. Assoc. J. 47: 515-519. Sturkie, P. D., 1947. Effects of hypothermia upon the specific gravity and proteins of the blood of chickens. Am. J. Physiol. 148: 610-613. Sturkie, P. D., 1950. Unpublished.
{Continued from page 230)
E. L. TerBush (B.S.A.—Cornell University) has been appointed Graduate Research Assistant in poultry genetics. H. F. Butlers (B.S.A.—University of Maine) is serving as Graduate Research Assistant in poultry nutrition. C. H. Moore (B.S.A.—Alabama Polytechnic Institute; M.S.—Kansas State College) was transferred from Graduate Research Assistant to Cooperative Agent with the Bureau of Animal Industry. He will assist Dr. Warren in the activities of the Regional Poultry Breeding Project. WASHINGTON NOTES
M. S. Mitchell has been appointed Teaching Assistant in the Department of Poultry Husbandry. He is a graduate of the University of Manitoba, Winnipeg, Canada and will proceed with degree work for the M.S., specializing in poultry genetics. T. Myint (Ph.D.—-Utah State College) has been appointed Research Associate in the Department of Poultry Husbandry, working on the project dealing with the development of microbiological techniques
in the analyses of amino acids and proteins. Dr. Myint is a native of Rangoon, Burma.
E. Sauter (B.S.—Washington State College) has been appointed Research Assistant in poultry products. He will investigate relationships between egg quality and culinary use of the egg, in cooperation with the Department of Foods and Nutrition in the College of Home Economics. OHIO NOTES
Appointments to graduate research assistantships in Poultry Science on projects of the Ohio Agricultural Experiment Station during the academic year 1950-51 are: P. L. Clayton (M.S., 1950, Ohio); L. S. Mercia (B.S., 1950, Vermont) and A. W. Jasper (B.S., 1950, Vermont) in economics and marketing; J. W. Marguand (B.S., 1948, Ohio) in turkey production; Charles McFarland (M.S., 1950, Ohio); D. W. Puffenberger (B.S., 1949, West Virginia) and C. R. Wettling (M.S., 1950, Ohio) in processing preservation of poultry products.
{Continued on page 266)
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News and Notes