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The Relationship of Vitamin K to the Incidence of Blood Spots in Eggs and Blood Prothrombin Time of Layers1
794
RESEARCH NOTES
REFERENCES Harms, R. H., and P. W. Waldroup, 1963. Length of laying cycle as influenced by dietary protein level. Poultry Sci. 42:1195-1197.
Waldroup, P. W., and R. H. Harms, 1962. Effect of grower diets on the performance of egg production type pullets. Florida Agric. Expt. Sta. Tech. Bull. 646.
THE RELATIONSHIP OF VITAMIN K TO THE INCIDENCE OF BLOOD SPOTS IN EGGS AND BLOOD PROTHROMBIN TIME OF LAYERS 1
Department of Poultry Science, Mississippi State University, State College, Mississippi (Received for publication February 20, 1964)
We are aware of only one report indicating that supplemental vitamin K decreases the incidence of blood spots in eggs (Berruti and Didrick, 1961). A recent report by Suater et al. (1963) indicates that supplemental alfalfa meal, a rich source of vitamin K, increases egg blood spotting. Waldroup and Harms (1962) added dicumarol, a vitamin K antagonist, to a layer ration and failed to significantly increase blood spotting of eggs. These workers also concluded that blood prothrombin time was not related to the incidence of blood spots in eggs. The present study was conducted to determine the effect of menadione sodium bisulfite complex (MSBC) and alfalfa meal on blood spotting of eggs and to determine if there is a relationship between blood prothrombin time and the incidence of blood spots in eggs. EXPERIMENTAL
White Leghorn layers were maintained in individual cages and allowed feed and water ad libitum. Three basal diets were fed with and without the addition of MSBC as shown in Table 1. Each of the test diets was fed to replicate groups of 17 layers. During the last 87 days of the 11 1 Mississippi Agricultural Experiment Journal Article No. 1194.
Station
TABLE 1.—Composition of the basal layer rations Ingredient
Yellow corn Soybean meal, 5 0 % protein Limestone Defluorinated rock phosphate, 14% P 29% Ca Sodium chloride Dehydrated alfalfa meal, 17% protein Dried whey, 6 5 % lactose Fish meal, 60% protein Premix 1 Total
Basal ration number 1
%
2
%
3
%
72.34 20.00 4.50
70.84 19.00 4.50
69.99 16.25 4.50
2.50 0.40
2.50 0.40
2.10 0.40
— — — 0.26
2.50
— — 0.26
2.50 2.00 2.00 0.26
100.00
100.00
100.00
1
Premix furnished the following vitamins per ton of feed: Vitamin A, 4,082,400 I.U.; vitamin D3, 1.0 million I.C.U.; riboflavin, 3 gms.; pantothenic acid, 6 gms.; niacin, 15.5 gms.; choline chloride, 340.2 gms.; vitamin B12, 4.5 mg.
months experimental period, 34 hens exhibiting a high egg blood spotting record, were removed from the original test groups and subdivided for further testing. One group was fed basal ration 1 and another group received the same ration supplemented with MSBC (20 gm./ton). During each test period, all eggs laid on two consecutive days each week were broken out and checked for blood spots. A glass plate mirror device was used which allowed full view of the eggs. Blood prothrombin determinations were made at the end of the test period. RESULTS AND DISCUSSION
Since egg production and feed utilization were not significantly affected by
Downloaded from http://ps.oxfordjournals.org/ at Michigan State University on February 18, 2015
ELBERT J. DAY AND ROBERT C. WOODY
795
RESEARCH NOTES T A B L E 2.—Effect
of supplemental menadione sodium bisulfite complex (MSBC) of blood spots in eggs and blood prothrombin time of layers
Ration1 Basal Basal Basal Basal Basal Basal
1 (corn-soy) 1 plus M S B C , 4 g m . / T 2 (corn-soy+alfalfa meal) 2 plus M S B C , 4 g m . / T 3 (corn-soy plus whey, alfalfa and fish meal) 3 plus M S B C 4 g m . / T
on the
incidence
N o . eggs broken out
N o . blood spots
% blood spots
Prothrombin time, sec.
1,985 1,968 1,874 1,819 1,882 1,983
80 136 96 96 98 107
4.0>2 6.9 b 5.1 a b 5.3»b 5.2»b 5.4»b
46.0 a 34.2 b ° 36.9b« 34.6 bc 41.4» b 33.4"
1
dietary treatment, these data are not presented. However, blood spotting of eggs and blood prothrombin times were affected by the type of ration fed, Tables 2 and 3. Supplemental MSBC increased the percentage of eggs containing blood spots as compared to each control ration; however, this increase was significant only with the groups fed basal ration 1. Alfalfa meal also tended to cause a higher incidence of blood spots but these differences were not statistically significant. Blood prothrombin time for the birds fed the simplified basal ration 1 was significantly lower than that of the other groups. The lowered prothrombin time along with a corresponding increase in blood spotting of eggs suggests a possible relationship between blood prothrombin time and egg blood spotting. A rise in T A B L E 3.—Effect of supplemental MSBC when fed to layers exhibiting a high incidence of egg blood spotting 87 day test period1 Ration* Basal 1 (corn-soy) Basal plus MSBC, 20 gm./T 1
No. eggs broken out
No. blood spots
% blood spots
Prothrombin time, sec.
148
24
16.2"!
46.0"
148
33
22.3 b
27.0 b
Birds in these groups were selected from the first six treatments in the previous study (Table 2) on the basis of their previous record of unusually high incidence of blood spotting. 2 See Table 1 for composition of the basal ration. 8 Any means within each column having different superscripts are significantly different at the 5% level of probability (Duncan, 1955).
blood prothrombin time may be beneficial, rather than harmful, in preventing blood spotting of eggs. Further tests are needed before definite conclusions can be made. Even if further data substantiate the hypothesis that decreasing clotting time is associated with increased blood spotting of eggs, the increase would probably not be due to more blood getting into the eggs, but to better clotting and hence better detection of the small amount of blood present. SUMMARY An increase in the incidence of blood spots in eggs was observed as a result of supplementing cage layer diets with MSBC or alfalfa meal. The differences obtained with alfalfa meal were not statistically significant. In two of four comparisons MSBC significantly increased blood spotting of eggs. Blood prothrombin time was decreased with supplemental MSBC or alfalfa meal, suggesting a possible inverse relationship between prothrombin time of layers and the incidence of egg blood spots. ACKNOWLEDGMENTS
This work was supported in part by a grant-in-aid from Heterochemical Corporation, Valley Stream, New York, New York.
Downloaded from http://ps.oxfordjournals.org/ at Michigan State University on February 18, 2015
See T a b l e 1 for t h e composition of t h e basal ration. A n y m e a n s within each column h a v i n g different superscripts are significantly different a t t h e 5 % level of probability ( D u n c a n , 1955). 2
796
RESEARCH NOTES
REFERENCES Berruti, R. and G. Dedrick, 1961. Evidence on the value of stabilized Heterogen " B " in reducing the incidence of blood spots in eggs. Unpublished data, Heterochemical Corporation, Valley Stream, New York, New York. Duncan, D. B., 1955. Multiple range and multiple
F tests. Biometrics, 11:1-42. Sauter, E. A., C. F. Petersen, C. E. Lampman and A. C. Wiese, 1963. Dietary influence upon blood spots incidence. Poultry Sci. 42:1304. Waldroup, P. W., and R. H. Harms, 1962. The influence of dicumarol on the incidence of blood spots. Poultry Sci. 41: 510-512.
ELBERT J. DAY, BEN C. DILWORTH AND P. N. DUA Department of Poultry Science, Mississippi Slate University, State College, Mississippi (Received for publication February 24, 1964)
Lowered blood prothrombin time of layers, as a result of adding menadione sodium bisulfite complex (MSBC) to the diet, has been associated with an increased incidence of blood spots in eggs (Day and Woody, 1964). In contrast, the addition of anti-coagulants to layer diets has reportedly prolonged blood clotting times without affecting the incidence of blood spots (Waldroup and Harms, 1962; Siddiqui and Fry, 1963). Both of the above research groups concluded that prothrombin time and the incidence of egg blood spots appear not to be correlated. However, a critical examination of the data presented shows that the anti-coagulants actually decreased the percentage of eggs containing blood spots. The preliminary experiment reported herein was conducted to study the effect of supplemental dicumarol, MSBC and sulfaquinoxaline (SQ) on: (1) the incidence and size of blood spots in eggs, (2) blood prothrombin times and, (3) egg production and feed utilization of cage layers. Each of five dietary treatments was fed to groups of 17 caged White Leghorn layers, replicated four times, for a test 1 Mississippi Agricultural Experiment Journal Article No. 1135.
Station
period of three months. The dietary additions were made to a simplified corn-soy basal. The incidence and size of blood spots were determined by collecting and breaking all eggs laid during three consecutive days, for three weeks of each month. Prothrombin determinations were made according to the method of Woody et al. (1963) using eight birds per treatment at the end of the test period. Statistical examinations were made using the analysis of variance (Snedecor, 1957) and multiple range test (Duncan, 1955). Dicumarol supplementation significantly reduced the percentage of eggs containing blood spots, Table 1. In addition, blood prothrombin time was significantly increased due to dicumarol supplementation. The single additions of MSBC or SQ had no significant effect on blood prothrombin time or incidence of egg blood spots. There was a slight tendency for blood spot incidence to be increased with either of these additives. In addition, blood prothrombin time tended to be prolonged due to SQ supplementation. The addition of both MSBC and SQ to the diet caused a significant increase in egg blood spot incidence without significantly affecting blood prothrombin time.
Downloaded from http://ps.oxfordjournals.org/ at Michigan State University on February 18, 2015
REDUCED INCIDENCE OF BLOOD SPOTS IN EGGS WITH DICUMAROL SUPPLEMENTATION 1
RESEARCH NOTES
REFERENCES Harms, R. H., and P. W. Waldroup, 1963. Length of laying cycle as influenced by dietary protein level. Poultry Sci. 42:1195-1197.
Waldroup, P. W., and R. H. Harms, 1962. Effect of grower diets on the performance of egg production type pullets. Florida Agric. Expt. Sta. Tech. Bull. 646.
THE RELATIONSHIP OF VITAMIN K TO THE INCIDENCE OF BLOOD SPOTS IN EGGS AND BLOOD PROTHROMBIN TIME OF LAYERS 1
Department of Poultry Science, Mississippi State University, State College, Mississippi (Received for publication February 20, 1964)
We are aware of only one report indicating that supplemental vitamin K decreases the incidence of blood spots in eggs (Berruti and Didrick, 1961). A recent report by Suater et al. (1963) indicates that supplemental alfalfa meal, a rich source of vitamin K, increases egg blood spotting. Waldroup and Harms (1962) added dicumarol, a vitamin K antagonist, to a layer ration and failed to significantly increase blood spotting of eggs. These workers also concluded that blood prothrombin time was not related to the incidence of blood spots in eggs. The present study was conducted to determine the effect of menadione sodium bisulfite complex (MSBC) and alfalfa meal on blood spotting of eggs and to determine if there is a relationship between blood prothrombin time and the incidence of blood spots in eggs. EXPERIMENTAL
White Leghorn layers were maintained in individual cages and allowed feed and water ad libitum. Three basal diets were fed with and without the addition of MSBC as shown in Table 1. Each of the test diets was fed to replicate groups of 17 layers. During the last 87 days of the 11 1 Mississippi Agricultural Experiment Journal Article No. 1194.
Station
TABLE 1.—Composition of the basal layer rations Ingredient
Yellow corn Soybean meal, 5 0 % protein Limestone Defluorinated rock phosphate, 14% P 29% Ca Sodium chloride Dehydrated alfalfa meal, 17% protein Dried whey, 6 5 % lactose Fish meal, 60% protein Premix 1 Total
Basal ration number 1
%
2
%
3
%
72.34 20.00 4.50
70.84 19.00 4.50
69.99 16.25 4.50
2.50 0.40
2.50 0.40
2.10 0.40
— — — 0.26
2.50
— — 0.26
2.50 2.00 2.00 0.26
100.00
100.00
100.00
1
Premix furnished the following vitamins per ton of feed: Vitamin A, 4,082,400 I.U.; vitamin D3, 1.0 million I.C.U.; riboflavin, 3 gms.; pantothenic acid, 6 gms.; niacin, 15.5 gms.; choline chloride, 340.2 gms.; vitamin B12, 4.5 mg.
months experimental period, 34 hens exhibiting a high egg blood spotting record, were removed from the original test groups and subdivided for further testing. One group was fed basal ration 1 and another group received the same ration supplemented with MSBC (20 gm./ton). During each test period, all eggs laid on two consecutive days each week were broken out and checked for blood spots. A glass plate mirror device was used which allowed full view of the eggs. Blood prothrombin determinations were made at the end of the test period. RESULTS AND DISCUSSION
Since egg production and feed utilization were not significantly affected by
Downloaded from http://ps.oxfordjournals.org/ at Michigan State University on February 18, 2015
ELBERT J. DAY AND ROBERT C. WOODY
795
RESEARCH NOTES T A B L E 2.—Effect
of supplemental menadione sodium bisulfite complex (MSBC) of blood spots in eggs and blood prothrombin time of layers
Ration1 Basal Basal Basal Basal Basal Basal
1 (corn-soy) 1 plus M S B C , 4 g m . / T 2 (corn-soy+alfalfa meal) 2 plus M S B C , 4 g m . / T 3 (corn-soy plus whey, alfalfa and fish meal) 3 plus M S B C 4 g m . / T
on the
incidence
N o . eggs broken out
N o . blood spots
% blood spots
Prothrombin time, sec.
1,985 1,968 1,874 1,819 1,882 1,983
80 136 96 96 98 107
4.0>2 6.9 b 5.1 a b 5.3»b 5.2»b 5.4»b
46.0 a 34.2 b ° 36.9b« 34.6 bc 41.4» b 33.4"
1
dietary treatment, these data are not presented. However, blood spotting of eggs and blood prothrombin times were affected by the type of ration fed, Tables 2 and 3. Supplemental MSBC increased the percentage of eggs containing blood spots as compared to each control ration; however, this increase was significant only with the groups fed basal ration 1. Alfalfa meal also tended to cause a higher incidence of blood spots but these differences were not statistically significant. Blood prothrombin time for the birds fed the simplified basal ration 1 was significantly lower than that of the other groups. The lowered prothrombin time along with a corresponding increase in blood spotting of eggs suggests a possible relationship between blood prothrombin time and egg blood spotting. A rise in T A B L E 3.—Effect of supplemental MSBC when fed to layers exhibiting a high incidence of egg blood spotting 87 day test period1 Ration* Basal 1 (corn-soy) Basal plus MSBC, 20 gm./T 1
No. eggs broken out
No. blood spots
% blood spots
Prothrombin time, sec.
148
24
16.2"!
46.0"
148
33
22.3 b
27.0 b
Birds in these groups were selected from the first six treatments in the previous study (Table 2) on the basis of their previous record of unusually high incidence of blood spotting. 2 See Table 1 for composition of the basal ration. 8 Any means within each column having different superscripts are significantly different at the 5% level of probability (Duncan, 1955).
blood prothrombin time may be beneficial, rather than harmful, in preventing blood spotting of eggs. Further tests are needed before definite conclusions can be made. Even if further data substantiate the hypothesis that decreasing clotting time is associated with increased blood spotting of eggs, the increase would probably not be due to more blood getting into the eggs, but to better clotting and hence better detection of the small amount of blood present. SUMMARY An increase in the incidence of blood spots in eggs was observed as a result of supplementing cage layer diets with MSBC or alfalfa meal. The differences obtained with alfalfa meal were not statistically significant. In two of four comparisons MSBC significantly increased blood spotting of eggs. Blood prothrombin time was decreased with supplemental MSBC or alfalfa meal, suggesting a possible inverse relationship between prothrombin time of layers and the incidence of egg blood spots. ACKNOWLEDGMENTS
This work was supported in part by a grant-in-aid from Heterochemical Corporation, Valley Stream, New York, New York.
Downloaded from http://ps.oxfordjournals.org/ at Michigan State University on February 18, 2015
See T a b l e 1 for t h e composition of t h e basal ration. A n y m e a n s within each column h a v i n g different superscripts are significantly different a t t h e 5 % level of probability ( D u n c a n , 1955). 2
796
RESEARCH NOTES
REFERENCES Berruti, R. and G. Dedrick, 1961. Evidence on the value of stabilized Heterogen " B " in reducing the incidence of blood spots in eggs. Unpublished data, Heterochemical Corporation, Valley Stream, New York, New York. Duncan, D. B., 1955. Multiple range and multiple
F tests. Biometrics, 11:1-42. Sauter, E. A., C. F. Petersen, C. E. Lampman and A. C. Wiese, 1963. Dietary influence upon blood spots incidence. Poultry Sci. 42:1304. Waldroup, P. W., and R. H. Harms, 1962. The influence of dicumarol on the incidence of blood spots. Poultry Sci. 41: 510-512.
ELBERT J. DAY, BEN C. DILWORTH AND P. N. DUA Department of Poultry Science, Mississippi Slate University, State College, Mississippi (Received for publication February 24, 1964)
Lowered blood prothrombin time of layers, as a result of adding menadione sodium bisulfite complex (MSBC) to the diet, has been associated with an increased incidence of blood spots in eggs (Day and Woody, 1964). In contrast, the addition of anti-coagulants to layer diets has reportedly prolonged blood clotting times without affecting the incidence of blood spots (Waldroup and Harms, 1962; Siddiqui and Fry, 1963). Both of the above research groups concluded that prothrombin time and the incidence of egg blood spots appear not to be correlated. However, a critical examination of the data presented shows that the anti-coagulants actually decreased the percentage of eggs containing blood spots. The preliminary experiment reported herein was conducted to study the effect of supplemental dicumarol, MSBC and sulfaquinoxaline (SQ) on: (1) the incidence and size of blood spots in eggs, (2) blood prothrombin times and, (3) egg production and feed utilization of cage layers. Each of five dietary treatments was fed to groups of 17 caged White Leghorn layers, replicated four times, for a test 1 Mississippi Agricultural Experiment Journal Article No. 1135.
Station
period of three months. The dietary additions were made to a simplified corn-soy basal. The incidence and size of blood spots were determined by collecting and breaking all eggs laid during three consecutive days, for three weeks of each month. Prothrombin determinations were made according to the method of Woody et al. (1963) using eight birds per treatment at the end of the test period. Statistical examinations were made using the analysis of variance (Snedecor, 1957) and multiple range test (Duncan, 1955). Dicumarol supplementation significantly reduced the percentage of eggs containing blood spots, Table 1. In addition, blood prothrombin time was significantly increased due to dicumarol supplementation. The single additions of MSBC or SQ had no significant effect on blood prothrombin time or incidence of egg blood spots. There was a slight tendency for blood spot incidence to be increased with either of these additives. In addition, blood prothrombin time tended to be prolonged due to SQ supplementation. The addition of both MSBC and SQ to the diet caused a significant increase in egg blood spot incidence without significantly affecting blood prothrombin time.
Downloaded from http://ps.oxfordjournals.org/ at Michigan State University on February 18, 2015
REDUCED INCIDENCE OF BLOOD SPOTS IN EGGS WITH DICUMAROL SUPPLEMENTATION 1