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The Relationship of Vitamin K to Mortality from Cecal Coccidiosis1
The Relationship of Vitamin K to Mortality from Cecal Coccidiosis1 R. L. TUGWELL, J. F. STEPHENS AND R. H. HARMS Poultry Department, University of Tennessee, Knoxville (Received for publication May 24, 1957)
H
EXPERIMENTAL PROCEDURE
Ten pens of males and ten pens of females (Lancaster X New Hampshire) were used in the first experiment. Twenty pens of Single Comb White Leghorn males were used in the second experiment. In either experiment each pen contained twenty, day-old birds. Five experimental treatments were used resulting in four replicates being used with each treatment. Eight pens of birds were fed vitamin K low basal diet as shown in Table 1. Four of these pens served as the positive control and were not inoculated with E. tenella. The diets of the other three experimental groups were formed by the 1
Published with the approval of Director, Tennessee Agricultural Experiment Station. 2 Klotogen F, trademark Abbott Laboratories, which contains by definition 6 3 % sodium bisulfite, U.S.P.
addition of one gram of MSB per ton of feed, twenty grams of MSB per ton of feed, and three percent of alfalfa leaf meal respectively to the basal diet. These three experimental groups and four pens of the basal fed birds were inoculated with sporulated oocyts of E. tenella. Each bird in the first experiment was inoculated with 50,000 oocysts at 23 days of age while each of those in the second experiment received 60,000 oocysts at 28 days of age. All chicks were reared in electrically heated battery brooders with raised screen-wire floors. Feed and water were given ad libitum. Water troughs were washed and fresh water supplied daily in an effort to limit the synthesis of vitamin K in the watering troughs. The individual body weight of all chicks was obtained at time of inoculation and again fourteen days post inoculation. Mortality records were kept throughout the experiments. However, only mortality reTABLE 1.—Composition of basal diet Ingredient Yellow Corn Soybean Oil Meal (50% protein) Defluorinated Phosphate 1 Ground Limestone Salt Vitamin Mix2 1
Amt. per 100 lbs. 65.7 31.0 1.0 1.2 0.5 0.6
Contains 34% calcium and 17% phosphorus. Contains per pound of feed: 0.9 mg. riboflavin, 2 mg. calcium pantothenate, 5 mg. niacin, 3 meg. vitamin Bi 2 ,250 mg. choline chloride, 1737 i.tr. vitamin A, 1050 I.C.U. vitamin D 3 , 0.08 gms. MnS0 4 , 2 mg., procaine penicillin, and 45 mg. arsanilic acid.
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2
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ARMS and Tugwell (1956) reported that mortality from cecal coccidiosis in chicks could be significantly increased by the addition of dicumarol to the diet. This increased mortality was attributed to prolonging blood clotting time, since the addition of one gram per ton of menadione sodium bisulfite2 (hereafter referred to as MSB) significantly reduced mortality when added to the diet containing dicumarol. In view of these findings experiments were designed to study the possibility of reducing mortality from cecal coccidiosis by the addition of MSB to a vitamin K low diet.
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R.
L. TUGWELL, J. F. STEPHENS AND R.
H.
HARMS
Downloaded from http://ps.oxfordjournals.org/ at NERL on April 12, 2015
peared to furnish adequate vitamin K to prevent excessive mortality from coccidiosis. Recently Otto et al. (1957) have also shown that vitamin K is necessary to prevent excessive mortality from coccidiosis. The addition of a source of vitamin K to the basal diet, in the second experiment, resulted in significantly reducing ing blood clotting time (Table 2). This reduction in blood clotting time may have been expected since vitamin K has been shown to be necessary for normal blood clotting (Dam, 1935). I t is of interest to note that significant differences in mortality and blood clotting time occurred beRESULTS AND DISCUSSION tween the same groups. This would further substantiate the fact that sufficient vitaMortality of chicks suffering from cecal min K must be present in the diet to give coccidiosis was significantly reduced by "normal" blood clotting time in order to addition of a source of vitamin K to the avoid excessive mortality in the presence basal diet (Table 2). The addition of of cecal coccidiosis. either 3 % alfalfa, one gram or twenty grams of MSB per ton of feed significantly Weight gains were obtained 14 days reduced mortality when birds were arti- post inoculation. No significant differences ficially inoculated with E. tenetta. in weight gains were obtained between In the first experiment mortality of inoculated groups indicating that addibirds receiving twenty grams of MSB per tion of vitamin K was not effective in imton of feed was significantly less than that proving growth rate when the chicks were obtained with chicks receiving one gram subjected to cecal coccidiosis. of MSB per ton of feed. This would indiSUMMARY cate that one gram of MSB per ton of feed was not adequate during an attack of cocMortality of chicks artificially inocucidiosis. However, no significant difference lated with Eimeria tenella was signifiin mortality could be detected between cantly decreased when a source of vitamin these groups in the second experiment. In K was added to the basal diet. In one of each experiment, 3 % alfalfa leaf meal ap- two experiments the addition of twenty grams of MSB per ton of feed significantly TABLE 2.—Effect of vitamin K on blood clotting time decreased mortality when compared to feeding one gram per ton. Three percent and mortality from coccidiosis of alfalfa leaf meal appeared to supply an ExperiExperiment #2 adequate amount of vitamin K to prevent ment #1 • excessive mortality from coccidiosis. (pertime (percent) cent; (seconds) The addition of a source of vitamin K 0.0 0.0 824 Basal Non-Inoc. to the basal diet employed in these tests, Basal Inoc. 36.7 44.6 751 1 gm. Menadione/ton-Inoc. 468 19.2] 2.5 resulted in significantly reducing blood 3% Alfalfa Leaf Meal-Inoc. 13.21 6.3 545 20 gms. Menadione/ton-Inoc. 8.2 428 clotting time. Mortality of chicks arti11.7 1 ficially infected with Eimeria tenella was * Means not grouped by lines are significantly different. suiting from cecal coccidiosis is included in this report; an additional five chicks died from other causes. In the second experiment, thirty birds were selected at random from each treatment group for blood clotting time determinations. Clotting times were measured by the capillary tube method with blood obtained by puncturing the wing vein. Statements of probability in this paper are based on analysis of variance according to Snedecor (1956) with significant treatment differences determined by Duncan's multiple range test (1955).
VITAMIN K AND COCCIDIOSIS
significantly decreased when the blood clotting time was reduced. Further investigation is needed to establish the vitamin K requirement in the presence of an attack of cecal coccidiosis. ACKNOWLEDGMENTS This study was supported in part by a grant-in-aid from Abbott Laboratories, North Chicago, Illinois. The B vitamins and vitamin A used in these studies were supplied through the courtesy of Charles F. Pfizer Co., Terre Haute, Indiana.
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REFERENCES Dam, H., 1935. The antihemorrhagic vitamin of the chick. Nature, 135: 652-653. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11:1-42. Harms, R. H., and R. L. Tugwell, 1956. The effect of experimentally induced prolonged blood clotting time on cecal coccidiosis of chicks. Poultry Sci. 35: 937-938. Otto, G. F., H. A. Jeske, D. V. Frost and H. S. Perdue, 1957. Menadione sodium bisulfite complex (Klotogen F) in caecal coccidiosis. Poultry Sci. 36: 1147. Snedecor, G. W., 1956. Statistical Methods. Fifth Edition, The Iowa State College Press, Ames, Iowa
TILL M. HUSTON, W. PERRY JOINER 3 AND JAMES L. CARMON4
Poultry Division, University of Georgia, Athens, Georgia (Received for publication May 24, 1957)
N
teridge et al. (1944) increased the environmental temperature of laying pullets by the use of artificial heat and insulation over a range of 37.8°F. to 59.9°F. and got no effect upon egg production. Hutt (1938) reported breed differences in susceptibility to extreme heat. White Leghorns withstood drastic increases in environmental temperatures better than did Rhode Island Reds or Barred Plymouth Rocks. Other workers have made similar observations. Fox (1951) demonstrated that White Leghorn hens withstood high temperatures better than Rhode Island Red hens. Warren (1939) secured data on egg size from localities in latitudes extending from 1 Journal Series Paper No. 41, College Experiment the Equator to as far north as Scotland. The results showed that atmospheric temStation, University of Georgia, Athens, Georgia. 2 Supported in part by a research grant from the perature was a very important factor in American Poultry and Hatchery Federation. determining the egg size of domestic fowl. 3 Present address: Poultry Department, Rutgers Bennion and Warren (1933), Skoglund University, New Brunswick, New Jersey. 4 Geneticist and Statistician, Animal Science Di- et al. (1951), and Hutchinson (1953) showed a sharp decline in egg weight when vision, University of Georgia. UMEROUS studies at various geographic locations over a period of several years indicate rather conclusively that high environmental temperature is one of the major factors influencing the decline in egg production during the summer months. Winchester (1940), Byerly et al. (1947), Wilson (1948), Ragab and Assem (1953) and others have shown that reproductive performance of domestic fowl is adversely affected by high environmental temperatures. Bruckner (1936) found that in a poultry house artificially heated to a mean temperature of 60°F., the health, body weight, and egg size of the birds were adversely affected. Gut-
Downloaded from http://ps.oxfordjournals.org/ at NERL on April 12, 2015
Breed Differences in Egg Production of Domestic Fowl Held at High Environmental Temperatures1,2
H
EXPERIMENTAL PROCEDURE
Ten pens of males and ten pens of females (Lancaster X New Hampshire) were used in the first experiment. Twenty pens of Single Comb White Leghorn males were used in the second experiment. In either experiment each pen contained twenty, day-old birds. Five experimental treatments were used resulting in four replicates being used with each treatment. Eight pens of birds were fed vitamin K low basal diet as shown in Table 1. Four of these pens served as the positive control and were not inoculated with E. tenella. The diets of the other three experimental groups were formed by the 1
Published with the approval of Director, Tennessee Agricultural Experiment Station. 2 Klotogen F, trademark Abbott Laboratories, which contains by definition 6 3 % sodium bisulfite, U.S.P.
addition of one gram of MSB per ton of feed, twenty grams of MSB per ton of feed, and three percent of alfalfa leaf meal respectively to the basal diet. These three experimental groups and four pens of the basal fed birds were inoculated with sporulated oocyts of E. tenella. Each bird in the first experiment was inoculated with 50,000 oocysts at 23 days of age while each of those in the second experiment received 60,000 oocysts at 28 days of age. All chicks were reared in electrically heated battery brooders with raised screen-wire floors. Feed and water were given ad libitum. Water troughs were washed and fresh water supplied daily in an effort to limit the synthesis of vitamin K in the watering troughs. The individual body weight of all chicks was obtained at time of inoculation and again fourteen days post inoculation. Mortality records were kept throughout the experiments. However, only mortality reTABLE 1.—Composition of basal diet Ingredient Yellow Corn Soybean Oil Meal (50% protein) Defluorinated Phosphate 1 Ground Limestone Salt Vitamin Mix2 1
Amt. per 100 lbs. 65.7 31.0 1.0 1.2 0.5 0.6
Contains 34% calcium and 17% phosphorus. Contains per pound of feed: 0.9 mg. riboflavin, 2 mg. calcium pantothenate, 5 mg. niacin, 3 meg. vitamin Bi 2 ,250 mg. choline chloride, 1737 i.tr. vitamin A, 1050 I.C.U. vitamin D 3 , 0.08 gms. MnS0 4 , 2 mg., procaine penicillin, and 45 mg. arsanilic acid.
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2
Downloaded from http://ps.oxfordjournals.org/ at NERL on April 12, 2015
ARMS and Tugwell (1956) reported that mortality from cecal coccidiosis in chicks could be significantly increased by the addition of dicumarol to the diet. This increased mortality was attributed to prolonging blood clotting time, since the addition of one gram per ton of menadione sodium bisulfite2 (hereafter referred to as MSB) significantly reduced mortality when added to the diet containing dicumarol. In view of these findings experiments were designed to study the possibility of reducing mortality from cecal coccidiosis by the addition of MSB to a vitamin K low diet.
1246
R.
L. TUGWELL, J. F. STEPHENS AND R.
H.
HARMS
Downloaded from http://ps.oxfordjournals.org/ at NERL on April 12, 2015
peared to furnish adequate vitamin K to prevent excessive mortality from coccidiosis. Recently Otto et al. (1957) have also shown that vitamin K is necessary to prevent excessive mortality from coccidiosis. The addition of a source of vitamin K to the basal diet, in the second experiment, resulted in significantly reducing ing blood clotting time (Table 2). This reduction in blood clotting time may have been expected since vitamin K has been shown to be necessary for normal blood clotting (Dam, 1935). I t is of interest to note that significant differences in mortality and blood clotting time occurred beRESULTS AND DISCUSSION tween the same groups. This would further substantiate the fact that sufficient vitaMortality of chicks suffering from cecal min K must be present in the diet to give coccidiosis was significantly reduced by "normal" blood clotting time in order to addition of a source of vitamin K to the avoid excessive mortality in the presence basal diet (Table 2). The addition of of cecal coccidiosis. either 3 % alfalfa, one gram or twenty grams of MSB per ton of feed significantly Weight gains were obtained 14 days reduced mortality when birds were arti- post inoculation. No significant differences ficially inoculated with E. tenetta. in weight gains were obtained between In the first experiment mortality of inoculated groups indicating that addibirds receiving twenty grams of MSB per tion of vitamin K was not effective in imton of feed was significantly less than that proving growth rate when the chicks were obtained with chicks receiving one gram subjected to cecal coccidiosis. of MSB per ton of feed. This would indiSUMMARY cate that one gram of MSB per ton of feed was not adequate during an attack of cocMortality of chicks artificially inocucidiosis. However, no significant difference lated with Eimeria tenella was signifiin mortality could be detected between cantly decreased when a source of vitamin these groups in the second experiment. In K was added to the basal diet. In one of each experiment, 3 % alfalfa leaf meal ap- two experiments the addition of twenty grams of MSB per ton of feed significantly TABLE 2.—Effect of vitamin K on blood clotting time decreased mortality when compared to feeding one gram per ton. Three percent and mortality from coccidiosis of alfalfa leaf meal appeared to supply an ExperiExperiment #2 adequate amount of vitamin K to prevent ment #1 • excessive mortality from coccidiosis. (pertime (percent) cent; (seconds) The addition of a source of vitamin K 0.0 0.0 824 Basal Non-Inoc. to the basal diet employed in these tests, Basal Inoc. 36.7 44.6 751 1 gm. Menadione/ton-Inoc. 468 19.2] 2.5 resulted in significantly reducing blood 3% Alfalfa Leaf Meal-Inoc. 13.21 6.3 545 20 gms. Menadione/ton-Inoc. 8.2 428 clotting time. Mortality of chicks arti11.7 1 ficially infected with Eimeria tenella was * Means not grouped by lines are significantly different. suiting from cecal coccidiosis is included in this report; an additional five chicks died from other causes. In the second experiment, thirty birds were selected at random from each treatment group for blood clotting time determinations. Clotting times were measured by the capillary tube method with blood obtained by puncturing the wing vein. Statements of probability in this paper are based on analysis of variance according to Snedecor (1956) with significant treatment differences determined by Duncan's multiple range test (1955).
VITAMIN K AND COCCIDIOSIS
significantly decreased when the blood clotting time was reduced. Further investigation is needed to establish the vitamin K requirement in the presence of an attack of cecal coccidiosis. ACKNOWLEDGMENTS This study was supported in part by a grant-in-aid from Abbott Laboratories, North Chicago, Illinois. The B vitamins and vitamin A used in these studies were supplied through the courtesy of Charles F. Pfizer Co., Terre Haute, Indiana.
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REFERENCES Dam, H., 1935. The antihemorrhagic vitamin of the chick. Nature, 135: 652-653. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11:1-42. Harms, R. H., and R. L. Tugwell, 1956. The effect of experimentally induced prolonged blood clotting time on cecal coccidiosis of chicks. Poultry Sci. 35: 937-938. Otto, G. F., H. A. Jeske, D. V. Frost and H. S. Perdue, 1957. Menadione sodium bisulfite complex (Klotogen F) in caecal coccidiosis. Poultry Sci. 36: 1147. Snedecor, G. W., 1956. Statistical Methods. Fifth Edition, The Iowa State College Press, Ames, Iowa
TILL M. HUSTON, W. PERRY JOINER 3 AND JAMES L. CARMON4
Poultry Division, University of Georgia, Athens, Georgia (Received for publication May 24, 1957)
N
teridge et al. (1944) increased the environmental temperature of laying pullets by the use of artificial heat and insulation over a range of 37.8°F. to 59.9°F. and got no effect upon egg production. Hutt (1938) reported breed differences in susceptibility to extreme heat. White Leghorns withstood drastic increases in environmental temperatures better than did Rhode Island Reds or Barred Plymouth Rocks. Other workers have made similar observations. Fox (1951) demonstrated that White Leghorn hens withstood high temperatures better than Rhode Island Red hens. Warren (1939) secured data on egg size from localities in latitudes extending from 1 Journal Series Paper No. 41, College Experiment the Equator to as far north as Scotland. The results showed that atmospheric temStation, University of Georgia, Athens, Georgia. 2 Supported in part by a research grant from the perature was a very important factor in American Poultry and Hatchery Federation. determining the egg size of domestic fowl. 3 Present address: Poultry Department, Rutgers Bennion and Warren (1933), Skoglund University, New Brunswick, New Jersey. 4 Geneticist and Statistician, Animal Science Di- et al. (1951), and Hutchinson (1953) showed a sharp decline in egg weight when vision, University of Georgia. UMEROUS studies at various geographic locations over a period of several years indicate rather conclusively that high environmental temperature is one of the major factors influencing the decline in egg production during the summer months. Winchester (1940), Byerly et al. (1947), Wilson (1948), Ragab and Assem (1953) and others have shown that reproductive performance of domestic fowl is adversely affected by high environmental temperatures. Bruckner (1936) found that in a poultry house artificially heated to a mean temperature of 60°F., the health, body weight, and egg size of the birds were adversely affected. Gut-
Downloaded from http://ps.oxfordjournals.org/ at NERL on April 12, 2015
Breed Differences in Egg Production of Domestic Fowl Held at High Environmental Temperatures1,2