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Hematocrit of Chickens; with Special Reference to Visceral Lymphomatosis
Hematocrit of Chickens; with Special Reference to Visceral Lymphomatosis KOJI SATO Department of Animal Physiology, Faculty of Agriculture, Nagoya University, Anzyo, Japan (Received for publication December 15, 1959)
R
EXPERIMENTAL
Cross-bred hens of White Leghorn X Rhode Island Red available at the Nagoya University Farm Experimental Station, were used for these experiments. Samples of blood were drawn from the wing vein with heparin being employed as the anticoagulant. The blood was introduced into a hematocrit tube which had been devised by Wintrobe, and this tube was centrifuged for 30 minutes at 3,000
rpm. (1,500 X G). Relative volumes of M and the erythrocytes (E), were determined by reading the values from a graduated scale affixed to the tube. The chickens used were clinically suspected of having visceral lymphomatosis from their general manifestations and this diagnosis was assured pathohistologically through the courtesy of the Kyushu Regional Laboratory, National Institute of Animal Health. Normal relative volumes of M and E were obtained from the laying hens and roosters which were considered to be healthy on the day of determination. The hemoglobin was determined colorimetrically as oxyhemoglobin and the number of leucocytes was enumerated by the Shaw method. RESULTS AND DISCUSSION
The data in sections A and B in Table 1 were obtained from chickens having visceral lymphomatosis; however, the chickens in section B did not show any enlargement of the liver. The results of control experiments with emaciated chickens displaying the same general manifestations are shown in section C. As the manifestations became worse, E in section A increased slightly (No. 6, 12, etc). It might be assumed that hemoconcentration took place; however, the mean relative volume of E did not differ significantly from those of normal chickens (Table 2). A significantly higher relative volume for M was found in chickens showing the enlargement of the liver, kidney, and spleen due to visceral lymphomatosis. In
1126
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ELATIVE volumes of blood cells and plasma have been determined after centrifugation in a hematocrit tube. In such a separation of cells and plasma, the platelets (thrombocytes) and leucocytes, being less dense than the erythrocytes, appear as a separate layer above the erythrocytes. The color of the separate layer is reddish gray in human and chickens. Wintrobe (1956) stated that thickness of the reddish gray layer which is a mixture of platelets (thrombocytes) and leucocytes (M), increase in cases of human leukemia, and is a useful guide to the diagnosis of leukemia. As the number of leucocytes in leukemic patients increases, the increase in the relative volume of M would be readily conceivable. The prominent increase in M was found in chicken granuloblastosis (Jungherr, 1952). The present paper is concerned with the observations of chicken hematocrit. An increase in the relative volume of M was observed in cases of visceral lymphomatosis in which there was an enlargement of the liver, kidney, and spleen, even though the number of leucocytes increased but little.
1127
HEMATOCRIT OF CHICKENS TABLE 1.—Hematocrit values in chickens having visceral lymphomatosis and other diseases. All were females except No. 8 and 11 Days before Chicken No. death (days)
Hematocrit
Body weight (kg.)
E.
M.
White Oxyhemo- blood cell count globin (xioy (g./dl.) mm.3)
(%)
7 1
1.75 1.34
20.0 33.2
3.0 3.8
5.7 11.2
3.5 5.1
7*
7 0
1.79 1.36
30.8 34.0
3.7 3.0
10.6 10.4
1.3 7.9
12
9 3
2.06 1.74
34.0 40.0
3.8 3.8
10.2
2.7
16
0
1.65
22.4
2.8
8.3
1.8
17
5
2.05
24.4
2.4
12.2
2.0
19*
1
1.38
36.0
3.0
14.2
—
23
24 15
1.53 1.35
33.2 31.0
2.6 3.0
10.3
1.1
A 28
1
1.35
33.0
6.0
—
3.5
30
80 46 15
1.25 1.44
29.0 27.0 22.2
2.6 2.8 3.0
12.5
35
3
1.55
32.0
3.0
12.8
—
37*
16 0
1.75 1.16
29.0 25.2
2.6 2.8
8.8
7.2
39*
0
1.39
40.6
3.8
15.6
3.8
40
18 1
1.55 1.25
28.6 26.5
3.0 2.1
10.0 9.7
1.5 3.3
43
4
1.10
28.0
2.2
—
—
53*
0
1.90
34.0
3.8
—
—
54*
0
1.70
32.0
5.6
—
—
41*
3 0
0.75
35.3 37.0
1.6 1.0
—
1.2 1.6
B
Visceral lymphomatosis with liver atrophy
—
—
Myelocytoma in liver
1.1
—
1.9
Myelocytoma in ovary
48.0
0.8
—
2.1
Myxosarcoma in intestine
1.50
23.6
1.8
—
3.9
CC14 intoxication
4
1.60
44.0
0.2
13.3
1.4
CHCls intoxication
0
0.15
30.0
1.8
—
—
Pullorum disease
3 0
0.71
11.8 8.6
52
7
1.50
31.0
4.0
63
5
1.10
28.2
64*
0
1.50
9
3
C 65* * Killed.
E
—
42*
11* (c?)
Visceral lymphomatosis with liver enlargement
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
(%)
6
Remarks
1128
K. SATO TABLE 1—(continued)
Days before Chicken No. death (days)
Hematocrit
Body weight (kg.)
E.
M.
(%)
(%)
White Oxyhemo- blood globin cell count (X10V (g./dl.) mm.3)
0
0.25
30.0
2.0
—
—
—
1.20
36.1
1.2
8.5
3.4
Blackhead
33
36
1.38
35.0
2.0
9.8
—
Coccidiosis
32
10
1.24
23.0
2.0
7.3
—
Infested with Ascarides
67
4
—
17.0
1.8
—
—
Infested with Cestodes
66* 8(cf)
TABLE
2 . --Mean
values of E% and M% of hematocrit Hematocrit E(%)
M(%)
Lymphomatosis
(24) '
30.3±1.07 3.30±0.02
Laying Hen (A) (B) (Q
(36) (28) (27)
31.7+1.6 28.8+1.2 26.1 + 0. 5
1.80±0.05 1.78±0.07 1.70±0.07
Roosters
(12)
35.9+0.5
1.68±0.06 M+ S.E.
)*=Number of determinations. )t=Number of chickens.
M occurred before the clinical manifestations of lymphomatosis, it might be possible to diagnose chickens having lymphomatosis which otherwise could not be distinguished clinically from any of the other normal chickens. To explore this possibility an experiment, the results of which are shown in Table 3, was undertaken. Two day-old chickens were injected intraperitoneally with liver homogenate prepared from lymphomatosis chicken No. 40. Either at 107 or at 136 days after inoculation, above 2.0% M was found in all the chickens. No. 53 could not rise from the floor 264 days after inoculation; however, an increase of the relative volume of M was not observed 29 to 89 days before this attack. The increase in the relative volume of M was found to have occurred only at measurement taken 128 days prior to death, a stage in which the disease could be diagnosed by clinical signs and symptoms. Two of the chickens in group A in Table 2 showing a 1.8% and 1.6% M, died after 30 days and 53 days respectively. Autopsy revealed that both chickens had visceral lymphomatosis with the enlargement of the liver. However, No. 30, a chicken in Table 1 (section A), while showing a relatively larger volume of M, 2.6%, was alive for 80 days after the first day of determination and during that time increased in body weight and laid
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
healthy chickens (Table 2), there was no difference among the groups in the mean relative volumes of M while those of E fluctuated by reflecting the difference in either sex, feeding, and or laying conditions. In the 103 healthy chickens in this experiment (Table 2), the relative volumes of M were in the range of from 0.9% to 2.7%. Only 4 chickens showed above 2.0% M, namely: 2.2, 2.4, 2.6, and 2.7%. The chicken showing 2.7%, of M manifested signs of visceral lymphomatosis 40 days after the day of determination (No. 37 in Table 1). However, the increase in the relative volume of M was not specific to visceral lymphomatosis as shown in No. 41 and 42 (section B) and No. 52 (section C). The liver of No. 52 was enlarged and showed massive infiltration of myelocytes and plasma cells through microscopic examination. If an increase in the relative volume of
Pullorum disease
HEMATOCRIT or CHICKENS
1129
TABLE 3.—Observations of chicken hematocrit after inoculation with liver homo%enate Liver homogenate obtained from chicken No. 40 in Table 1 was injected intraperitoneally to nine chickens 2 days old on May 15,1958. Two of them died of coccidiosis within 30 days after inoculation. Days after inoculationi Chicken No.
S1 51
B.W.kg. He. E.% M%
61
(July)
107 (Aug.)
0.38 31.2 1.9
0.40 23.1 2.8
136 (Sep.)
175 (Oct.)
204 (Nov.)
235 (Dec.)
Remarks Aug. 22, '58, killed. Visceral lymphomatosis
0.40
1.00 28.6 1.6
1.35 31.8 2.2
1.61 32.9 1.9
555 3
B W - He. E. M.
0.42 26.0 1.8
0.88 31.2 1.2
1.10 27.8 2.2
1.40 34.4 1.0
„ B.W. He. E. M.
0.42 30.3 2.0
0.65 28.0 3.0
0.95 31.4 1.8
1.22 34.0 1.4
1.50 34.0 1.4
1.75 31.0 1.2
May 29, '59, died. Myelocytoma
E. M.
0.31 29.7 1.3
0.80 30.2 1.2
0.90 26.0 2.6
1.35 31.2 1.6
1.75 33.0 1.0
2.02 31.0 1.0
June 2, '59, killed Myxosarcoma
B.W. He. E. M.
0.40 30.3 2.0
0.80 28.0 3.0
1.25 31.4 1.8
1.50 34.0 1.4
2.10 34.0 1.4
2.25 31.0 1.2
Oct. 17, '59, died Egg concretion
fA. 64
79 /y
B
-W-
He.
eggs from May to June. The enlargement of the liver could not be ascertained by palpation. When it finally died the relative volume of M was 3.0% and autopsy showed a moderate liver enlargement, thus this chicken may be said to have been a chronic case. According to the Wintrobe's description, the relative volume of M depends upon the number and kinds of leucocytes and the number of platelets. The increase in the relative volume of M in human leukemia keeps pace with the increase in the number of leucocytes, but it is not always a quantitative increase when the number of leucocytes increases over 30,000/mm3. As obviously indicated in Table 1, in chickens M had no correlation to the number of leucocytes. The kinds of leucocytes were scrutinized in blood films stained with Giemsa. The distribution pattern of the leucocytes did not differ very much from the normal pattern. In No. 28 and No. 54 the number of thrombocytes increased. It
2.20 30.2 1.4
Nov. '58, ran away
might be supposed that the largest relative volume of M shown in Table 1 was caused by an increase in the thrombocytes; however, in all the cases except No. 28 and No. 54 an increase in the thrombocytes was not found. When M was microscopically examined, erythrocytes were also found with the thrombocytes and leucocytes (Fig. 1). In addition to Wintrobe's description, two other possible reasons for the cause of an increase of the relative volume of M may be considered. The first is that there is an increase in the number of erythrocytes having the same density as the leucocytes upon which the relative volume of M depends, thus increasing the relative volume of M. The second concerns the compactness of M.M became a compact mass above the erythrocytes layer in which the erythrocytes kept their fluidity when sampled by using a pipette. The relative volume of M was changed by the addition of egg albumin, or increased by the ligation
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
B W - He. E. M.
63
2.20 30.4 1.2
Feb. 3,'59, killed. Visceral lymphomatosis
53 3,3
1130
K.
»
SATO
^pr %
<
•
c V
#
-V
. 1
•
•
«
ft
•
mf % m
•
9%
FIG. 1. Microscopic view of the reddish gray layer in the hematocrit. It is composed of leucocytes, platelets and a few erythrocytes. Preparation was obtained from No. .S4 and stained with Giemsa.
of the cystic duct of a normal chicken; therefore, it could be supposed that unknown factor or factors which determine the extent of the compactness of M or cohesive property of the leucocytes to the erythrocytes might appear in the blood of lymphomatosis chickens when their liver was severely invaded. This also may explain the relatively larger volume of M in No. 52. Further studies concerning the relative volume of M will be required. ACKNOWLEDGMENT
The author gratefully acknowledges the cooperation and advice of all staff members of the Nagoya University Farm Experimental Station, Anzyo. Dr. K. Iwashina, Kyushu Regional Laboratory, National Institute of Animal Health, and Dr. T. Chiba, Research Institute of the Environmental Medicine, Nagoya University, also gave valuable suggestions for which I am very appreciative.
SUMMARY (1) The relative volume of a mixed layer (M) of the thrombocytes and leucocytes and of the erythrocytes was determined in the blood of chicken by the hematocrit-method. (2) While the relative volume of E in visceral lymphomatosis chickens did not differ from t h a t in healthy chickens, the relative volume of M was greater in visceral lymphomatosis chickens with enlargement of the liver; however, such alteration was not considered specific for this disease. (3) The possible causes for the increase in the relative volume of M are discussed. REFERENCES Jungherr, E., 1952. Diseases of Poultry, edited by H. E. Biester and L. H. Schwarte, Chapter 18, Avian leucosis complex. Iowa State College Press, Ames, Iowa. Wintrobe, M. W., 1956. Clinical Hematology. Lea and Febiger, Philadelphia.
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
ift
• / • «C
R
EXPERIMENTAL
Cross-bred hens of White Leghorn X Rhode Island Red available at the Nagoya University Farm Experimental Station, were used for these experiments. Samples of blood were drawn from the wing vein with heparin being employed as the anticoagulant. The blood was introduced into a hematocrit tube which had been devised by Wintrobe, and this tube was centrifuged for 30 minutes at 3,000
rpm. (1,500 X G). Relative volumes of M and the erythrocytes (E), were determined by reading the values from a graduated scale affixed to the tube. The chickens used were clinically suspected of having visceral lymphomatosis from their general manifestations and this diagnosis was assured pathohistologically through the courtesy of the Kyushu Regional Laboratory, National Institute of Animal Health. Normal relative volumes of M and E were obtained from the laying hens and roosters which were considered to be healthy on the day of determination. The hemoglobin was determined colorimetrically as oxyhemoglobin and the number of leucocytes was enumerated by the Shaw method. RESULTS AND DISCUSSION
The data in sections A and B in Table 1 were obtained from chickens having visceral lymphomatosis; however, the chickens in section B did not show any enlargement of the liver. The results of control experiments with emaciated chickens displaying the same general manifestations are shown in section C. As the manifestations became worse, E in section A increased slightly (No. 6, 12, etc). It might be assumed that hemoconcentration took place; however, the mean relative volume of E did not differ significantly from those of normal chickens (Table 2). A significantly higher relative volume for M was found in chickens showing the enlargement of the liver, kidney, and spleen due to visceral lymphomatosis. In
1126
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
ELATIVE volumes of blood cells and plasma have been determined after centrifugation in a hematocrit tube. In such a separation of cells and plasma, the platelets (thrombocytes) and leucocytes, being less dense than the erythrocytes, appear as a separate layer above the erythrocytes. The color of the separate layer is reddish gray in human and chickens. Wintrobe (1956) stated that thickness of the reddish gray layer which is a mixture of platelets (thrombocytes) and leucocytes (M), increase in cases of human leukemia, and is a useful guide to the diagnosis of leukemia. As the number of leucocytes in leukemic patients increases, the increase in the relative volume of M would be readily conceivable. The prominent increase in M was found in chicken granuloblastosis (Jungherr, 1952). The present paper is concerned with the observations of chicken hematocrit. An increase in the relative volume of M was observed in cases of visceral lymphomatosis in which there was an enlargement of the liver, kidney, and spleen, even though the number of leucocytes increased but little.
1127
HEMATOCRIT OF CHICKENS TABLE 1.—Hematocrit values in chickens having visceral lymphomatosis and other diseases. All were females except No. 8 and 11 Days before Chicken No. death (days)
Hematocrit
Body weight (kg.)
E.
M.
White Oxyhemo- blood cell count globin (xioy (g./dl.) mm.3)
(%)
7 1
1.75 1.34
20.0 33.2
3.0 3.8
5.7 11.2
3.5 5.1
7*
7 0
1.79 1.36
30.8 34.0
3.7 3.0
10.6 10.4
1.3 7.9
12
9 3
2.06 1.74
34.0 40.0
3.8 3.8
10.2
2.7
16
0
1.65
22.4
2.8
8.3
1.8
17
5
2.05
24.4
2.4
12.2
2.0
19*
1
1.38
36.0
3.0
14.2
—
23
24 15
1.53 1.35
33.2 31.0
2.6 3.0
10.3
1.1
A 28
1
1.35
33.0
6.0
—
3.5
30
80 46 15
1.25 1.44
29.0 27.0 22.2
2.6 2.8 3.0
12.5
35
3
1.55
32.0
3.0
12.8
—
37*
16 0
1.75 1.16
29.0 25.2
2.6 2.8
8.8
7.2
39*
0
1.39
40.6
3.8
15.6
3.8
40
18 1
1.55 1.25
28.6 26.5
3.0 2.1
10.0 9.7
1.5 3.3
43
4
1.10
28.0
2.2
—
—
53*
0
1.90
34.0
3.8
—
—
54*
0
1.70
32.0
5.6
—
—
41*
3 0
0.75
35.3 37.0
1.6 1.0
—
1.2 1.6
B
Visceral lymphomatosis with liver atrophy
—
—
Myelocytoma in liver
1.1
—
1.9
Myelocytoma in ovary
48.0
0.8
—
2.1
Myxosarcoma in intestine
1.50
23.6
1.8
—
3.9
CC14 intoxication
4
1.60
44.0
0.2
13.3
1.4
CHCls intoxication
0
0.15
30.0
1.8
—
—
Pullorum disease
3 0
0.71
11.8 8.6
52
7
1.50
31.0
4.0
63
5
1.10
28.2
64*
0
1.50
9
3
C 65* * Killed.
E
—
42*
11* (c?)
Visceral lymphomatosis with liver enlargement
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
(%)
6
Remarks
1128
K. SATO TABLE 1—(continued)
Days before Chicken No. death (days)
Hematocrit
Body weight (kg.)
E.
M.
(%)
(%)
White Oxyhemo- blood globin cell count (X10V (g./dl.) mm.3)
0
0.25
30.0
2.0
—
—
—
1.20
36.1
1.2
8.5
3.4
Blackhead
33
36
1.38
35.0
2.0
9.8
—
Coccidiosis
32
10
1.24
23.0
2.0
7.3
—
Infested with Ascarides
67
4
—
17.0
1.8
—
—
Infested with Cestodes
66* 8(cf)
TABLE
2 . --Mean
values of E% and M% of hematocrit Hematocrit E(%)
M(%)
Lymphomatosis
(24) '
30.3±1.07 3.30±0.02
Laying Hen (A) (B) (Q
(36) (28) (27)
31.7+1.6 28.8+1.2 26.1 + 0. 5
1.80±0.05 1.78±0.07 1.70±0.07
Roosters
(12)
35.9+0.5
1.68±0.06 M+ S.E.
)*=Number of determinations. )t=Number of chickens.
M occurred before the clinical manifestations of lymphomatosis, it might be possible to diagnose chickens having lymphomatosis which otherwise could not be distinguished clinically from any of the other normal chickens. To explore this possibility an experiment, the results of which are shown in Table 3, was undertaken. Two day-old chickens were injected intraperitoneally with liver homogenate prepared from lymphomatosis chicken No. 40. Either at 107 or at 136 days after inoculation, above 2.0% M was found in all the chickens. No. 53 could not rise from the floor 264 days after inoculation; however, an increase of the relative volume of M was not observed 29 to 89 days before this attack. The increase in the relative volume of M was found to have occurred only at measurement taken 128 days prior to death, a stage in which the disease could be diagnosed by clinical signs and symptoms. Two of the chickens in group A in Table 2 showing a 1.8% and 1.6% M, died after 30 days and 53 days respectively. Autopsy revealed that both chickens had visceral lymphomatosis with the enlargement of the liver. However, No. 30, a chicken in Table 1 (section A), while showing a relatively larger volume of M, 2.6%, was alive for 80 days after the first day of determination and during that time increased in body weight and laid
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
healthy chickens (Table 2), there was no difference among the groups in the mean relative volumes of M while those of E fluctuated by reflecting the difference in either sex, feeding, and or laying conditions. In the 103 healthy chickens in this experiment (Table 2), the relative volumes of M were in the range of from 0.9% to 2.7%. Only 4 chickens showed above 2.0% M, namely: 2.2, 2.4, 2.6, and 2.7%. The chicken showing 2.7%, of M manifested signs of visceral lymphomatosis 40 days after the day of determination (No. 37 in Table 1). However, the increase in the relative volume of M was not specific to visceral lymphomatosis as shown in No. 41 and 42 (section B) and No. 52 (section C). The liver of No. 52 was enlarged and showed massive infiltration of myelocytes and plasma cells through microscopic examination. If an increase in the relative volume of
Pullorum disease
HEMATOCRIT or CHICKENS
1129
TABLE 3.—Observations of chicken hematocrit after inoculation with liver homo%enate Liver homogenate obtained from chicken No. 40 in Table 1 was injected intraperitoneally to nine chickens 2 days old on May 15,1958. Two of them died of coccidiosis within 30 days after inoculation. Days after inoculationi Chicken No.
S1 51
B.W.kg. He. E.% M%
61
(July)
107 (Aug.)
0.38 31.2 1.9
0.40 23.1 2.8
136 (Sep.)
175 (Oct.)
204 (Nov.)
235 (Dec.)
Remarks Aug. 22, '58, killed. Visceral lymphomatosis
0.40
1.00 28.6 1.6
1.35 31.8 2.2
1.61 32.9 1.9
555 3
B W - He. E. M.
0.42 26.0 1.8
0.88 31.2 1.2
1.10 27.8 2.2
1.40 34.4 1.0
„ B.W. He. E. M.
0.42 30.3 2.0
0.65 28.0 3.0
0.95 31.4 1.8
1.22 34.0 1.4
1.50 34.0 1.4
1.75 31.0 1.2
May 29, '59, died. Myelocytoma
E. M.
0.31 29.7 1.3
0.80 30.2 1.2
0.90 26.0 2.6
1.35 31.2 1.6
1.75 33.0 1.0
2.02 31.0 1.0
June 2, '59, killed Myxosarcoma
B.W. He. E. M.
0.40 30.3 2.0
0.80 28.0 3.0
1.25 31.4 1.8
1.50 34.0 1.4
2.10 34.0 1.4
2.25 31.0 1.2
Oct. 17, '59, died Egg concretion
fA. 64
79 /y
B
-W-
He.
eggs from May to June. The enlargement of the liver could not be ascertained by palpation. When it finally died the relative volume of M was 3.0% and autopsy showed a moderate liver enlargement, thus this chicken may be said to have been a chronic case. According to the Wintrobe's description, the relative volume of M depends upon the number and kinds of leucocytes and the number of platelets. The increase in the relative volume of M in human leukemia keeps pace with the increase in the number of leucocytes, but it is not always a quantitative increase when the number of leucocytes increases over 30,000/mm3. As obviously indicated in Table 1, in chickens M had no correlation to the number of leucocytes. The kinds of leucocytes were scrutinized in blood films stained with Giemsa. The distribution pattern of the leucocytes did not differ very much from the normal pattern. In No. 28 and No. 54 the number of thrombocytes increased. It
2.20 30.2 1.4
Nov. '58, ran away
might be supposed that the largest relative volume of M shown in Table 1 was caused by an increase in the thrombocytes; however, in all the cases except No. 28 and No. 54 an increase in the thrombocytes was not found. When M was microscopically examined, erythrocytes were also found with the thrombocytes and leucocytes (Fig. 1). In addition to Wintrobe's description, two other possible reasons for the cause of an increase of the relative volume of M may be considered. The first is that there is an increase in the number of erythrocytes having the same density as the leucocytes upon which the relative volume of M depends, thus increasing the relative volume of M. The second concerns the compactness of M.M became a compact mass above the erythrocytes layer in which the erythrocytes kept their fluidity when sampled by using a pipette. The relative volume of M was changed by the addition of egg albumin, or increased by the ligation
Downloaded from http://ps.oxfordjournals.org/ at East Carolina University on June 27, 2015
B W - He. E. M.
63
2.20 30.4 1.2
Feb. 3,'59, killed. Visceral lymphomatosis
53 3,3
1130
K.
»
SATO
^pr %
<
•
c V
#
-V
. 1
•
•
«
ft
•
mf % m
•
9%
FIG. 1. Microscopic view of the reddish gray layer in the hematocrit. It is composed of leucocytes, platelets and a few erythrocytes. Preparation was obtained from No. .S4 and stained with Giemsa.
of the cystic duct of a normal chicken; therefore, it could be supposed that unknown factor or factors which determine the extent of the compactness of M or cohesive property of the leucocytes to the erythrocytes might appear in the blood of lymphomatosis chickens when their liver was severely invaded. This also may explain the relatively larger volume of M in No. 52. Further studies concerning the relative volume of M will be required. ACKNOWLEDGMENT
The author gratefully acknowledges the cooperation and advice of all staff members of the Nagoya University Farm Experimental Station, Anzyo. Dr. K. Iwashina, Kyushu Regional Laboratory, National Institute of Animal Health, and Dr. T. Chiba, Research Institute of the Environmental Medicine, Nagoya University, also gave valuable suggestions for which I am very appreciative.
SUMMARY (1) The relative volume of a mixed layer (M) of the thrombocytes and leucocytes and of the erythrocytes was determined in the blood of chicken by the hematocrit-method. (2) While the relative volume of E in visceral lymphomatosis chickens did not differ from t h a t in healthy chickens, the relative volume of M was greater in visceral lymphomatosis chickens with enlargement of the liver; however, such alteration was not considered specific for this disease. (3) The possible causes for the increase in the relative volume of M are discussed. REFERENCES Jungherr, E., 1952. Diseases of Poultry, edited by H. E. Biester and L. H. Schwarte, Chapter 18, Avian leucosis complex. Iowa State College Press, Ames, Iowa. Wintrobe, M. W., 1956. Clinical Hematology. Lea and Febiger, Philadelphia.
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