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The Influence of Dicumarol on the Incidence of Blood Spots in Eggs1
510
C. R. SADLER AND B. GLICK
body titer to Vibrio fetus bacterin of 1:64 or greater while only 50% of the birds in the "low" line had a titer of this magnitude. The data suggest that bursa size may influence antibody production. REFERENCES
The Influence of Dicumarol on the Incidence of Blood Spots in Eggs 1 P. W. WALDROUP AND R. H. HARMS Florida Agricultural Experiment Stations, Gainesville, Florida (Received for publication June 19, 1961)
T
HE occurrence of blood spots in eggs is a problem which causes great losses in marketable eggs, as well as unfavorable reaction from consumers. It is commonly assumed that from § to 6 percent of chicken eggs randomly selected contain blood spots. The exact cause of blood spots in eggs has not been determined, however, the occurrence of blood spots has been associated with certain physiological functions. Nalbandov and Card (1944) found that blood spots were due to the rupturing of blood vessels in the follicle wall. It was later reported that blood spots were due to hemorrhages that occur before ovulation (Taylor, 1949). It has recently been suggested that the addition of stabilized vitamin K to a hen feed will greatly reduce the percentage of blood spots occurring in eggs (Berruti and Dedrick, 1961). These workers postulated that the vitamin K was functioning in preventing hemorrhages of blood vessels inside the yolk sac and follicle. 1
Florida Agricultural Journal Series No. 1288.
Experiment
Stations,
Since dicumarol has been shown to counteract vitamin K, and significantly prolong the blood clotting time of chickens (Quick and Stefanini, 1948; Harms and Tarver, 1957), it was thought that feeding hens a level of dicumarol sufficient to greatly prolong blood clotting time might cause an increase in the incidence of blood spots by inducing hemorrhages of blood vessels inside the yolk sac and follicle. EXPERIMENTAL
Four experiments involving 130 hens, both light weight commercial egg production type and heavy meat type hens, have been conducted to determine the influence of feeding dicumarol on the incidence of blood spots in eggs. In all experiments, the hens were kept in individual cages and allowed the experimental feeds and water ad libitum The hens were randomly assigned to the various groups and records were kept for a period of one week to determine individual variation in rate of production and incidence of blood spots. The experimental feeds were fed to the hens
Downloaded from http://ps.oxfordjournals.org/ at Bibliothekssystem der Universitaet Giessen on June 14, 2015
Chang, T. S., M. S. Rheins and A. R. Winter, 1957. The significance of the bursa of Fabricius in antibody production in chickens. 1. Age of chicken.
Poultry Sci. 36; 735-739. Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Glick, B., 1956. Normal growth of the bursa of Fabricius in chickens. Poultry Sci. 35: 843-851. Glick, B., T. S. Chang and R. G. Jaap, 1956. The bursa of Fabricius and antibody production. Poultry Sci. 35: 224-225. Hepler, O. E., 1952. Manual of Clinical Laboratory Methods. Charles C Thomas, Springfield, 111. Snedecor, G. W., 1956. Statistical Methods. The Iowa State College Press, Ames, Iowa.
DlCUMAROL AND BLOOD SPOTS TABLE 1.—Composition of the basal diet Lbs./cwt.
Yellow corn Soybean oil meal (50% protein) Stabilized animal fat Alfalfa meal (17% protein) Ground limestone Defluorinated phosphate (34% Ca + 1 8 % P) Iodized salt Vitamin premix*
64.7 21.4 3.5 3.0 4.2
Rock, Rhode Island Red, and New Hampshire) were fed on each of the two experimental diets in experiment 3. RESULTS AND DISCUSSION
Feeding any of the three levels of dicumarol used in this experiment resulted 2.3 in significantly prolonging blood clotting 0.4 time (Table 2). It should be pointed out 0.5 that this increase in clotting time re* Supplied per pound of feed: 2,000 I.U. vitamin sulted even though the diet contained 3 A, 700 I.C.U. vitamin D 3 , 6 meg. vitamin Bi 2 , 2 mg. percent dehydrated alfalfa meal. Howriboflavin, 4 mg. calcium pantothenate, 6 mg. niacin, 227 mg. choline chloride, 2.5 I.U. vitamin E, and ever, no supplemental source of vitamin 80 mg. MnS0 4 . K was added to the feed. The incidence of blood spots was not until the hens receiving the dicumarol in their feed had died or practically ceased increased by the feeding of dicumarol production. This ranged from two to (Table 3). This was true even with the group of heavy breed hens which produced three weeks for each experiment. All eggs laid by each hen in each experi- eggs with a high incidence of blood spots ment were broken out on a glass plate prior to being placed on the experimental arranged with mirrors so a full view of feeds. It was found that birds producing the egg could be inspected for the presence eggs with blood spots during the preexperimental period laid more eggs with of blood spots. The composition of the basal diet is blood spots during the experimental shown in Table 1. Levels of 0, 500, 1,000, period than did birds laying eggs without and 1,500 mg. of dicumarol per pound blood spots during the pre-experimental were fed in experiment 1 in order to period. However, the size of blood spots determine the level of dicumarol neces- in eggs produced was not affected by the sary to increase blood clotting time with addition of dicumarol to the feed. laying hens. Two groups, each containing These results indicate that normal five egg production type hens, were fed blood clotting time is not essential in on each of the experimental diets. At the order to avoid the production of eggs with end of three days, individual blood clot- blood spots. These findings are in disting times were measured by the capillary agreement with results obtained by tube method, using blood obtained by puncturing the brachial vein. TABLE 2.—Blood clotting time of commercial egg production type pullets fed various levels of Since 500 mg. of dicumarol per pound dicumarol for a three day period of feed was sufficient to prolong clotting time in experiment 1, this was the only Number Average clotting Dicumarol pullets/ level used in succeeding experiments. (mg./lb.) time (min.-sec)1 treatment Groups of five egg production type 6' 18" 10 0 pullets were used in experiments 2 and 4. 10 41' 56" 500 Four groups were used per treatment in 1,000 10 51' 26" 10 42' 44" 1,500 experiment 2, and three groups per treatment in experiment 4. Five pullets of 1 All levels of dicumarol significantly increased three different breeds (White Plymouth clotting time when compared to the basal group.
Downloaded from http://ps.oxfordjournals.org/ at Bibliothekssystem der Universitaet Giessen on June 14, 2015
Ingredient
511
512
P. W. WALDROUP AND R. H. HARMS
TABLE 3.—Number of blood spots in broken-out eggs from pullets receiving diets containing dicumarol Exp. 1* Presence or absence of dicumarol
+
Exp. 2"
Exp. 3»
Exp.. 4»
All expts.
No. eggs
No. spots
No. eggs
No. spots
No. eggs
No. spots
No. eggs
No. spots
No. eggs
No. spots
% spots
34
0
77
2
56
32
186
8
353
42
11.8
58
0
70
5
30
14
124
0
282
19
6.7
a
Berruti and Dedrick (1961). No explanation can be given for this difference.
Laboratories, North Chicago, Illinois. REFERENCES
SUMMARY
Four experiments were conducted to determine the influence of feeding dicumarol upon incidence of blood spots in eggs. No significant increase was found in the incidence of blood spots in eggs from hens receiving the dicumarol-supplemented feed. The feeding of a diet supplemented with 500, 1,000, or 1,500 mg. per pound of dicumarol significantly increased the blood clotting time of laying hens. It is concluded that blood clotting time is not related to incidence of blood spots in eggs. ACKNOWLEDGMENT
This investigation was supported in part by a grant-in-aid from Abbott
Berruti, R., and G. Dedrick, 1961. Evidence on the value of stabilized heterogen K in reducing the incidence of blood spots in egg. Unpublished data. Heterochemical Corp., Valley Stream, New York. Harms, R. H., and F. R. Tarver, Jr., 1957. The influence of dicumarol upon blood clotting time and blood loss of young chickens. Poultry Sci. 36: 76-79. Nalbandov, A. V., and L. E. Card, 1944. The problem of blood clots and meat spots in chicken eggs. Poultry Sci. 23: 170-180. Quick, A. J., and M. Stefanini, 1948. Experimentally induced changes in the prothrombin level of the blood. IV. The relation of vitamin K deficiency to the intensity of dicumarol action and to the effect of excess vitamin assay. J. Biol. Chem. 175: 945-952. Snedecor, G. W., 1957. Statistical Methods. The Iowa State College Press, Ames, Iowa. Taylor, L. W., 1949. Fertility and Hatchability of Chicken and Turkey Eggs. John Wiley and Sons, New York.
NEWS AND NOTES (Continued from page 467) methodology; (3) professional statisticians who wish to keep informed about advanced specialized theory and methods; (4) prospective candidates for graduate degrees in statistics; and (5) graduate students in other fields who desire supporting work in statistics. The 1962 session, unlike the previous sessions, will last nine weeks instead of six, and will run concurrently with the regular summer session of the host institution. This will provide for nine semester hours of credit which may be applied as residence credit
at any one of the cooperating institutions in partial fulfillment of the requirement for a graduate degree. The program may be entered at any session and consecutive courses will follow in successive years, i.e., the program is unitary and sequential. A special feature, introduced this year, provides for completion of six hours of theory in nine weeks, thereby accelerating the program. The courses in statistics to be offered in 1962 at Oklahoma State University are: 4 courses in Methods, including Basic Statistical Methods, Experi-
{Continued on page 545)
Downloaded from http://ps.oxfordjournals.org/ at Bibliothekssystem der Universitaet Giessen on June 14, 2015
Light weight commercial egg production type pullets used in these experiments. Meat type (White Plymouth Rock, Rhode Island Red, and New Hampshire) pullets used in this experiment. b
C. R. SADLER AND B. GLICK
body titer to Vibrio fetus bacterin of 1:64 or greater while only 50% of the birds in the "low" line had a titer of this magnitude. The data suggest that bursa size may influence antibody production. REFERENCES
The Influence of Dicumarol on the Incidence of Blood Spots in Eggs 1 P. W. WALDROUP AND R. H. HARMS Florida Agricultural Experiment Stations, Gainesville, Florida (Received for publication June 19, 1961)
T
HE occurrence of blood spots in eggs is a problem which causes great losses in marketable eggs, as well as unfavorable reaction from consumers. It is commonly assumed that from § to 6 percent of chicken eggs randomly selected contain blood spots. The exact cause of blood spots in eggs has not been determined, however, the occurrence of blood spots has been associated with certain physiological functions. Nalbandov and Card (1944) found that blood spots were due to the rupturing of blood vessels in the follicle wall. It was later reported that blood spots were due to hemorrhages that occur before ovulation (Taylor, 1949). It has recently been suggested that the addition of stabilized vitamin K to a hen feed will greatly reduce the percentage of blood spots occurring in eggs (Berruti and Dedrick, 1961). These workers postulated that the vitamin K was functioning in preventing hemorrhages of blood vessels inside the yolk sac and follicle. 1
Florida Agricultural Journal Series No. 1288.
Experiment
Stations,
Since dicumarol has been shown to counteract vitamin K, and significantly prolong the blood clotting time of chickens (Quick and Stefanini, 1948; Harms and Tarver, 1957), it was thought that feeding hens a level of dicumarol sufficient to greatly prolong blood clotting time might cause an increase in the incidence of blood spots by inducing hemorrhages of blood vessels inside the yolk sac and follicle. EXPERIMENTAL
Four experiments involving 130 hens, both light weight commercial egg production type and heavy meat type hens, have been conducted to determine the influence of feeding dicumarol on the incidence of blood spots in eggs. In all experiments, the hens were kept in individual cages and allowed the experimental feeds and water ad libitum The hens were randomly assigned to the various groups and records were kept for a period of one week to determine individual variation in rate of production and incidence of blood spots. The experimental feeds were fed to the hens
Downloaded from http://ps.oxfordjournals.org/ at Bibliothekssystem der Universitaet Giessen on June 14, 2015
Chang, T. S., M. S. Rheins and A. R. Winter, 1957. The significance of the bursa of Fabricius in antibody production in chickens. 1. Age of chicken.
Poultry Sci. 36; 735-739. Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Glick, B., 1956. Normal growth of the bursa of Fabricius in chickens. Poultry Sci. 35: 843-851. Glick, B., T. S. Chang and R. G. Jaap, 1956. The bursa of Fabricius and antibody production. Poultry Sci. 35: 224-225. Hepler, O. E., 1952. Manual of Clinical Laboratory Methods. Charles C Thomas, Springfield, 111. Snedecor, G. W., 1956. Statistical Methods. The Iowa State College Press, Ames, Iowa.
DlCUMAROL AND BLOOD SPOTS TABLE 1.—Composition of the basal diet Lbs./cwt.
Yellow corn Soybean oil meal (50% protein) Stabilized animal fat Alfalfa meal (17% protein) Ground limestone Defluorinated phosphate (34% Ca + 1 8 % P) Iodized salt Vitamin premix*
64.7 21.4 3.5 3.0 4.2
Rock, Rhode Island Red, and New Hampshire) were fed on each of the two experimental diets in experiment 3. RESULTS AND DISCUSSION
Feeding any of the three levels of dicumarol used in this experiment resulted 2.3 in significantly prolonging blood clotting 0.4 time (Table 2). It should be pointed out 0.5 that this increase in clotting time re* Supplied per pound of feed: 2,000 I.U. vitamin sulted even though the diet contained 3 A, 700 I.C.U. vitamin D 3 , 6 meg. vitamin Bi 2 , 2 mg. percent dehydrated alfalfa meal. Howriboflavin, 4 mg. calcium pantothenate, 6 mg. niacin, 227 mg. choline chloride, 2.5 I.U. vitamin E, and ever, no supplemental source of vitamin 80 mg. MnS0 4 . K was added to the feed. The incidence of blood spots was not until the hens receiving the dicumarol in their feed had died or practically ceased increased by the feeding of dicumarol production. This ranged from two to (Table 3). This was true even with the group of heavy breed hens which produced three weeks for each experiment. All eggs laid by each hen in each experi- eggs with a high incidence of blood spots ment were broken out on a glass plate prior to being placed on the experimental arranged with mirrors so a full view of feeds. It was found that birds producing the egg could be inspected for the presence eggs with blood spots during the preexperimental period laid more eggs with of blood spots. The composition of the basal diet is blood spots during the experimental shown in Table 1. Levels of 0, 500, 1,000, period than did birds laying eggs without and 1,500 mg. of dicumarol per pound blood spots during the pre-experimental were fed in experiment 1 in order to period. However, the size of blood spots determine the level of dicumarol neces- in eggs produced was not affected by the sary to increase blood clotting time with addition of dicumarol to the feed. laying hens. Two groups, each containing These results indicate that normal five egg production type hens, were fed blood clotting time is not essential in on each of the experimental diets. At the order to avoid the production of eggs with end of three days, individual blood clot- blood spots. These findings are in disting times were measured by the capillary agreement with results obtained by tube method, using blood obtained by puncturing the brachial vein. TABLE 2.—Blood clotting time of commercial egg production type pullets fed various levels of Since 500 mg. of dicumarol per pound dicumarol for a three day period of feed was sufficient to prolong clotting time in experiment 1, this was the only Number Average clotting Dicumarol pullets/ level used in succeeding experiments. (mg./lb.) time (min.-sec)1 treatment Groups of five egg production type 6' 18" 10 0 pullets were used in experiments 2 and 4. 10 41' 56" 500 Four groups were used per treatment in 1,000 10 51' 26" 10 42' 44" 1,500 experiment 2, and three groups per treatment in experiment 4. Five pullets of 1 All levels of dicumarol significantly increased three different breeds (White Plymouth clotting time when compared to the basal group.
Downloaded from http://ps.oxfordjournals.org/ at Bibliothekssystem der Universitaet Giessen on June 14, 2015
Ingredient
511
512
P. W. WALDROUP AND R. H. HARMS
TABLE 3.—Number of blood spots in broken-out eggs from pullets receiving diets containing dicumarol Exp. 1* Presence or absence of dicumarol
+
Exp. 2"
Exp. 3»
Exp.. 4»
All expts.
No. eggs
No. spots
No. eggs
No. spots
No. eggs
No. spots
No. eggs
No. spots
No. eggs
No. spots
% spots
34
0
77
2
56
32
186
8
353
42
11.8
58
0
70
5
30
14
124
0
282
19
6.7
a
Berruti and Dedrick (1961). No explanation can be given for this difference.
Laboratories, North Chicago, Illinois. REFERENCES
SUMMARY
Four experiments were conducted to determine the influence of feeding dicumarol upon incidence of blood spots in eggs. No significant increase was found in the incidence of blood spots in eggs from hens receiving the dicumarol-supplemented feed. The feeding of a diet supplemented with 500, 1,000, or 1,500 mg. per pound of dicumarol significantly increased the blood clotting time of laying hens. It is concluded that blood clotting time is not related to incidence of blood spots in eggs. ACKNOWLEDGMENT
This investigation was supported in part by a grant-in-aid from Abbott
Berruti, R., and G. Dedrick, 1961. Evidence on the value of stabilized heterogen K in reducing the incidence of blood spots in egg. Unpublished data. Heterochemical Corp., Valley Stream, New York. Harms, R. H., and F. R. Tarver, Jr., 1957. The influence of dicumarol upon blood clotting time and blood loss of young chickens. Poultry Sci. 36: 76-79. Nalbandov, A. V., and L. E. Card, 1944. The problem of blood clots and meat spots in chicken eggs. Poultry Sci. 23: 170-180. Quick, A. J., and M. Stefanini, 1948. Experimentally induced changes in the prothrombin level of the blood. IV. The relation of vitamin K deficiency to the intensity of dicumarol action and to the effect of excess vitamin assay. J. Biol. Chem. 175: 945-952. Snedecor, G. W., 1957. Statistical Methods. The Iowa State College Press, Ames, Iowa. Taylor, L. W., 1949. Fertility and Hatchability of Chicken and Turkey Eggs. John Wiley and Sons, New York.
NEWS AND NOTES (Continued from page 467) methodology; (3) professional statisticians who wish to keep informed about advanced specialized theory and methods; (4) prospective candidates for graduate degrees in statistics; and (5) graduate students in other fields who desire supporting work in statistics. The 1962 session, unlike the previous sessions, will last nine weeks instead of six, and will run concurrently with the regular summer session of the host institution. This will provide for nine semester hours of credit which may be applied as residence credit
at any one of the cooperating institutions in partial fulfillment of the requirement for a graduate degree. The program may be entered at any session and consecutive courses will follow in successive years, i.e., the program is unitary and sequential. A special feature, introduced this year, provides for completion of six hours of theory in nine weeks, thereby accelerating the program. The courses in statistics to be offered in 1962 at Oklahoma State University are: 4 courses in Methods, including Basic Statistical Methods, Experi-
{Continued on page 545)
Downloaded from http://ps.oxfordjournals.org/ at Bibliothekssystem der Universitaet Giessen on June 14, 2015
Light weight commercial egg production type pullets used in these experiments. Meat type (White Plymouth Rock, Rhode Island Red, and New Hampshire) pullets used in this experiment. b