- Email: [email protected]
The Relative Differential Leucocyte Count of the Newly Hatched Chick1
ESTROGEN AND/OR ANDROGEN FOR LAYERS
Phillips, R. E., 1959. Endocrine mechanism of the failure of pintails (Anas acuta) to reproduce in captivity. Ph.D. Thesis, Cornell University. Schoorl, P., and H. Y. Boersma, 1962. Research on the quality of the eggshell. A new method of determination. Proc. 12th Wld's. Poultry Congr. p. 432-435. Steel and Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc., p. 72 and 99-146. Sturkie, P. D., 1965. Avian Physiology, second edition. Comstock Publishing Associates, Ithaca, New York, p. 447-501 and 568-585. Turner, C. W., 1948. Feeding estrogen (dianisylhexene) to laying hens. Poultry Sci. 27: 593-600. Urist, M. R., 1959. The effect of calcium deprivation upon the blood, adrenal cortex, ovary and skeleton in domestic fowl. Recent Progr. Horm. Res. 15:455-481. Zlatkis, A., B. Zak and A. J. Boyle, 1953. A new method for direct determination of serum cholesterol. J. Lab. Clinical Med. 41: 486-492.
The Relative Differential Leucocyte Count of the Newly Hatched Chick1 R. R. BURTON AND J. S. HARRISON Department of Animal Physiology, University of California, Davis, California 95616 (Received for publication August 14, 1968)
R
ECENTLY our laboratory made - some routine differential leucocyte counts on newly hatched chicks—0 to 31 days of age. The very young chicks had a relative lymphopenia, heterophilia, and basophilia compared with the hematological findings of adult chickens. Lucas and Jamroz (1961) also have reported similar leucocytic findings in the circulating blood of newly hatched chicks. Their research covered the first 8 days posthatch with considerable emphasis on the very early ages (1 hour to 42 hours posthatch). They reported an essentially normal proportion of the different types of white cells 1
This study was supported by NASA (Grant NGR-05-004-008).
in the circulating blood of 6-day old chicks. However, their data were limited to only four chicks per age group and were not statistically summarized. Therefore, since we had comparable data and since our findings were not identical to those of Lucas and Jamroz (1961), the information is reported herein. METHODS AND RESULTS
Differential leucocyte counts were determined by methods reported previously by Burton and Guion (1968). The blood was taken via heart puncture and heparin was used as the anticoagulant. The findings are summarized in Table 1. The lymphocytes and heterophils comprise approximately 90% of the total
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 27, 2015
Kessler, G., and M. Wolfman, 1964. An automated procedure for the simultaneous determination of calcium and phosphorus. Clin. Chem. 10: 686703. Kessler, G., and H. Lederer, 1965. Technicon symposia. Automation in analytical chemistry. Flurometric measurements of triglycerides and phospholipids, p. 341-344. Lorenz, F. W., and G. H. Bachman, 1947. Lipemia and fat deposition in response to oral administration of synthetic estrogens. Poultry Sci. 26: 419431. Marsh, W. H., B. Fingerhut and E. Kirsch, 1959. Adaptation of an alkaline phosphatase method for automatic colorimetric analysis. Clin. Chem. 5: 119-126. Martini, L., and W. F. Ganong, 1966. Neuroendocrinology, Volume 1, Academic Press, New York and London, p. 613-654. Nalbandov, A. V., 1964. Reproductive Physiology. W. M. Freeman and Co., San Francisco, Second edition, p. 144-148.
451
452
R. R. BURTON AND J. S. HARRISON TABLE 1.—The relative differential leucocyte counts in chicks of various ages (group means + standard errors) n1
Lymphocytes
Heterophils
Monocytes
Eosinophils
Basophils
0
11
15.9 + 2.52
72.4 + 3.28
2.5 + 0.79
1.1 + 0.44
8.1 + 2.69
2 3 4
9 9 4
26.7 + 4.34 38.7 + 5.60 24.0 + 2.12
62.8 + 5.61 52.7 + 5.70 64.8+1.50
2.1 + 0.61 1.6+0.78 0.75 + 0.48
1.1±0.45 0.67 + 0.33 2.8+1.03
7.3 + 1.12 6.4+1.13 7.8+1.38
6
3
47.7 + 4.06
44.3 + 2.73
1.3 + 0.33
2.7+1.33
4.0+1.00
8 9
8 5
48.3 + 4.40 32.4+4.86
50.0+4.60 64.2 + 5.33
0.25 + 0.16 1.8+0.49
0+ 0 4.0 + 0.24
1.5 + 0.46 1.2 + 0.49
11
6
58.3 + 8.60
26.8 + 7.27
3.7+1.17
3.7 + 0.95
7.3+1.31
13 16
14 4
50.2 + 4.70 64.0 + 0.58
41.8 + 4.40 30.8 + 0.85
2.9+1.10 2.8 + 0.25
0.57 + 0.20 0+0
4.5 + 1.10 2.5 + 0.29
20 31
11 7
68.6+3.30 69.4+6.05
26.7 + 2.83 25.1 + 6.19
1.7 + 0.43 2.0+0.72
0.64 + 0.24 0.43 + 0.30
2.3 + 0.68 2.9+1.18
1
Number of chicks per age group.
leucocytes in the circulating blood of chickens at all ages. However, the relationship of the lymphocytes to the heterophils rapidly changes during the first three weeks of life. Therefore, the lymphocyte :heterophil ratio—the percentage of lymphocytes divided by the percentage of heterophils—was used as a mathematical criterion of "hematological maturity." This ratio was mathematically related to the age of the chick and the following rectilinear equation was determined (Figure 1) : L : H = 0.16+0.116t
(1)
Where: L : H = lymphocyte:heterophil ratio; t = age of the chick in days (0 to 20 days); and, r (correlation coefficient) = 0.88; P<0.01 The lymphocyte :heterophil ratios of 2.5 and 2.8 were calculated for adult males and females respectively from the data reported by Lucas and Jamroz (1961; page 216). From equation (1) it appears
that "hematological maturity" occurs in the chick at approximately 20 days of age. Lucas and Jamroz (1961) report the near absence of monocytes in chicks less than 8 days of age which was not our finding (Table 1). Also of interest was the high percentage of basophils found in the young chicks—generally through 13 days of age. DISCUSSION
The dangers inherent in using relative leucocyte counts as opposed to total counts in estimating an animal's hematological status have been amply stated by Schalm (1965); however, Lucas and Jamroz (1961) have mathematically demonstrated that total leucocyte counts in the chicken have little statistical significance. Therefore, the following conclusions are based upon the chick's relative leucocyte count as reported herein. The young chick's leucocyte differential is characterized as high in heterophils and basophils and low in lymphocytes. This indicates a hematological defense mechanism oriented specifically to produce an
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 27, 2015
Age (days)
D I F F E R E N T I A L LEUCOCYTE
453
COUNT
3.02.8
o = ADULT MALES
2.6
D = ADULT FEMALES
2.4
• = DATA HEREIN
2.2
X
a. otr
1.8 l.6|L:H = 0.16 + 0.1161
1.4 1.2 LJ h>-
o o X
a.
r = 0.88,
P<0.0l
1.0 0.8 0.6 0.40.20.0
8
10 12 14 16 18 20 22 24 26 28 30
ADULTS
AGE (DAYS); t FIG. 1. The relationship of the lymphocyte: heterophil ratio to the age of the chick. The adult ratios were calculated from data of Lucas and Jamroz (1961; page 216).
inflammatory response—both heterophils and basophils in mammals are involved in the initial stages of inflammation. This ability at hatch to initiate an immediate inflammatory response is important since, probably, maternal passive immunity is not present in the oviparous chick. However, with the rapid increase in the circulating lymphocyte, it appears t h a t soon the chick has the capacity for antibody production. I n summary, the chick develops its "hematological m a t u r i t y " at approxi-
mately 20 days of age. At this time the entire relative leucocytic spectrum approximates adult values. REFERENCES Burton, R. R., and C. Guion, 1968. The differential leucocyte blood count: Its precision and individuality in the chicken. Poultry Sci. 47: 19451949. Lucas, A. M., and C. Jamroz, 1961. Atlas of Avian Hematology, Agri. Monog. 25. Wash., U. S. Dept. Agric. Schalm, O. W., 1965. Veterinary Hematology, 2nd Ed., Philadelphia: Lea and Febiger.
NEWS AND NOTES 'from page 443) Dr. George Jaap, W.P.S.A. Treasurer and Assistant Secretary, reported on the meeting of the Executive Committee which was held in Paris,
France, in February. The U.S.A. Branch has decided to submit an invitation to the World's Poultry Science Association
{Continued on page 474)
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 27, 2015
UJ
2.0
Phillips, R. E., 1959. Endocrine mechanism of the failure of pintails (Anas acuta) to reproduce in captivity. Ph.D. Thesis, Cornell University. Schoorl, P., and H. Y. Boersma, 1962. Research on the quality of the eggshell. A new method of determination. Proc. 12th Wld's. Poultry Congr. p. 432-435. Steel and Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc., p. 72 and 99-146. Sturkie, P. D., 1965. Avian Physiology, second edition. Comstock Publishing Associates, Ithaca, New York, p. 447-501 and 568-585. Turner, C. W., 1948. Feeding estrogen (dianisylhexene) to laying hens. Poultry Sci. 27: 593-600. Urist, M. R., 1959. The effect of calcium deprivation upon the blood, adrenal cortex, ovary and skeleton in domestic fowl. Recent Progr. Horm. Res. 15:455-481. Zlatkis, A., B. Zak and A. J. Boyle, 1953. A new method for direct determination of serum cholesterol. J. Lab. Clinical Med. 41: 486-492.
The Relative Differential Leucocyte Count of the Newly Hatched Chick1 R. R. BURTON AND J. S. HARRISON Department of Animal Physiology, University of California, Davis, California 95616 (Received for publication August 14, 1968)
R
ECENTLY our laboratory made - some routine differential leucocyte counts on newly hatched chicks—0 to 31 days of age. The very young chicks had a relative lymphopenia, heterophilia, and basophilia compared with the hematological findings of adult chickens. Lucas and Jamroz (1961) also have reported similar leucocytic findings in the circulating blood of newly hatched chicks. Their research covered the first 8 days posthatch with considerable emphasis on the very early ages (1 hour to 42 hours posthatch). They reported an essentially normal proportion of the different types of white cells 1
This study was supported by NASA (Grant NGR-05-004-008).
in the circulating blood of 6-day old chicks. However, their data were limited to only four chicks per age group and were not statistically summarized. Therefore, since we had comparable data and since our findings were not identical to those of Lucas and Jamroz (1961), the information is reported herein. METHODS AND RESULTS
Differential leucocyte counts were determined by methods reported previously by Burton and Guion (1968). The blood was taken via heart puncture and heparin was used as the anticoagulant. The findings are summarized in Table 1. The lymphocytes and heterophils comprise approximately 90% of the total
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 27, 2015
Kessler, G., and M. Wolfman, 1964. An automated procedure for the simultaneous determination of calcium and phosphorus. Clin. Chem. 10: 686703. Kessler, G., and H. Lederer, 1965. Technicon symposia. Automation in analytical chemistry. Flurometric measurements of triglycerides and phospholipids, p. 341-344. Lorenz, F. W., and G. H. Bachman, 1947. Lipemia and fat deposition in response to oral administration of synthetic estrogens. Poultry Sci. 26: 419431. Marsh, W. H., B. Fingerhut and E. Kirsch, 1959. Adaptation of an alkaline phosphatase method for automatic colorimetric analysis. Clin. Chem. 5: 119-126. Martini, L., and W. F. Ganong, 1966. Neuroendocrinology, Volume 1, Academic Press, New York and London, p. 613-654. Nalbandov, A. V., 1964. Reproductive Physiology. W. M. Freeman and Co., San Francisco, Second edition, p. 144-148.
451
452
R. R. BURTON AND J. S. HARRISON TABLE 1.—The relative differential leucocyte counts in chicks of various ages (group means + standard errors) n1
Lymphocytes
Heterophils
Monocytes
Eosinophils
Basophils
0
11
15.9 + 2.52
72.4 + 3.28
2.5 + 0.79
1.1 + 0.44
8.1 + 2.69
2 3 4
9 9 4
26.7 + 4.34 38.7 + 5.60 24.0 + 2.12
62.8 + 5.61 52.7 + 5.70 64.8+1.50
2.1 + 0.61 1.6+0.78 0.75 + 0.48
1.1±0.45 0.67 + 0.33 2.8+1.03
7.3 + 1.12 6.4+1.13 7.8+1.38
6
3
47.7 + 4.06
44.3 + 2.73
1.3 + 0.33
2.7+1.33
4.0+1.00
8 9
8 5
48.3 + 4.40 32.4+4.86
50.0+4.60 64.2 + 5.33
0.25 + 0.16 1.8+0.49
0+ 0 4.0 + 0.24
1.5 + 0.46 1.2 + 0.49
11
6
58.3 + 8.60
26.8 + 7.27
3.7+1.17
3.7 + 0.95
7.3+1.31
13 16
14 4
50.2 + 4.70 64.0 + 0.58
41.8 + 4.40 30.8 + 0.85
2.9+1.10 2.8 + 0.25
0.57 + 0.20 0+0
4.5 + 1.10 2.5 + 0.29
20 31
11 7
68.6+3.30 69.4+6.05
26.7 + 2.83 25.1 + 6.19
1.7 + 0.43 2.0+0.72
0.64 + 0.24 0.43 + 0.30
2.3 + 0.68 2.9+1.18
1
Number of chicks per age group.
leucocytes in the circulating blood of chickens at all ages. However, the relationship of the lymphocytes to the heterophils rapidly changes during the first three weeks of life. Therefore, the lymphocyte :heterophil ratio—the percentage of lymphocytes divided by the percentage of heterophils—was used as a mathematical criterion of "hematological maturity." This ratio was mathematically related to the age of the chick and the following rectilinear equation was determined (Figure 1) : L : H = 0.16+0.116t
(1)
Where: L : H = lymphocyte:heterophil ratio; t = age of the chick in days (0 to 20 days); and, r (correlation coefficient) = 0.88; P<0.01 The lymphocyte :heterophil ratios of 2.5 and 2.8 were calculated for adult males and females respectively from the data reported by Lucas and Jamroz (1961; page 216). From equation (1) it appears
that "hematological maturity" occurs in the chick at approximately 20 days of age. Lucas and Jamroz (1961) report the near absence of monocytes in chicks less than 8 days of age which was not our finding (Table 1). Also of interest was the high percentage of basophils found in the young chicks—generally through 13 days of age. DISCUSSION
The dangers inherent in using relative leucocyte counts as opposed to total counts in estimating an animal's hematological status have been amply stated by Schalm (1965); however, Lucas and Jamroz (1961) have mathematically demonstrated that total leucocyte counts in the chicken have little statistical significance. Therefore, the following conclusions are based upon the chick's relative leucocyte count as reported herein. The young chick's leucocyte differential is characterized as high in heterophils and basophils and low in lymphocytes. This indicates a hematological defense mechanism oriented specifically to produce an
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 27, 2015
Age (days)
D I F F E R E N T I A L LEUCOCYTE
453
COUNT
3.02.8
o = ADULT MALES
2.6
D = ADULT FEMALES
2.4
• = DATA HEREIN
2.2
X
a. otr
1.8 l.6|L:H = 0.16 + 0.1161
1.4 1.2 LJ h>-
o o X
a.
r = 0.88,
P<0.0l
1.0 0.8 0.6 0.40.20.0
8
10 12 14 16 18 20 22 24 26 28 30
ADULTS
AGE (DAYS); t FIG. 1. The relationship of the lymphocyte: heterophil ratio to the age of the chick. The adult ratios were calculated from data of Lucas and Jamroz (1961; page 216).
inflammatory response—both heterophils and basophils in mammals are involved in the initial stages of inflammation. This ability at hatch to initiate an immediate inflammatory response is important since, probably, maternal passive immunity is not present in the oviparous chick. However, with the rapid increase in the circulating lymphocyte, it appears t h a t soon the chick has the capacity for antibody production. I n summary, the chick develops its "hematological m a t u r i t y " at approxi-
mately 20 days of age. At this time the entire relative leucocytic spectrum approximates adult values. REFERENCES Burton, R. R., and C. Guion, 1968. The differential leucocyte blood count: Its precision and individuality in the chicken. Poultry Sci. 47: 19451949. Lucas, A. M., and C. Jamroz, 1961. Atlas of Avian Hematology, Agri. Monog. 25. Wash., U. S. Dept. Agric. Schalm, O. W., 1965. Veterinary Hematology, 2nd Ed., Philadelphia: Lea and Febiger.
NEWS AND NOTES 'from page 443) Dr. George Jaap, W.P.S.A. Treasurer and Assistant Secretary, reported on the meeting of the Executive Committee which was held in Paris,
France, in February. The U.S.A. Branch has decided to submit an invitation to the World's Poultry Science Association
{Continued on page 474)
Downloaded from http://ps.oxfordjournals.org/ at Florida International University on May 27, 2015
UJ
2.0