Prophylactic Efficacy of Nifursol Against Different Levels of Exposure to Histomoniasis in Turkeys 4 to 9 Weeks of Age1'2 T. W. SULLIVAN,3 R. J. MITCHELL 8 AND O. D. GRACE4 Departments of Poultry Science and Veterinary Science, University of Nebraska, Lincoln, Nebraska 68503 (Received for publication March 4, 1972)
POULTRY SCIENCE 51: 1956-1959,
INTRODUCTION
N
IFURSOL or 3,5-dinitrosalicylic acid, S-nitrofurylidene hydrazide is a new drug currently under investigation and development for the prevention of histomoniasis (blackhead) in poultry. Vatne et al. (1967, 1969) determined the prophylactic activity of nifursol against histomoniasis mortality and morbidity in poults. In this study poults received either a single oral inoculum of embryonated Heterakis gallinarium ova or were reared on soil contaminated continuously by droppings from H. gallinarium infected chickens in wire cages above the lots. Poults receiving a single infective dose were protected from blackhead 1 Published as paper number 3351, Journal Series, Nebraska Agricultural Experiment Station. 2 From a thesis submitted by R. J. Mitchell in partial fulfillment of the requirements for the M.S. degree. 3 Department of Poultry Science; R. J. Mitchell's present address: Rocco Inc., Harrisburg, Virginia 22801. 4 Department of Veterinary Science.
1972
mortality by feeding 0.004 through 0.2 percent nifursol. The lowest effective level was 0.00125 percent, which allowed survival of 44.5 percent of poults. All non-medicated control birds died of histomoniasis. In poults receiving continuous exposure, as previously described, 0.005 percent of nifursol in feed prevented all blackhead mortality in two experiments, and allowed only 5.3 percent mortality in a third. Sullivan et al. (1968) presented data concerning the prophylactic efficacy of nifursol against histomoniasis in turkeys four to 18 weeks of age. Nifursol levels of 0.00125 to 0.01 percent in feed prevented histomoniasis mortality and morbidity. The present study was conducted to determine the prophylactic efficacy of different levels of nifursol against graded levels of exposure to Histomoniasis in tur&eys iour to nine weeks of age. EXPERIMENTAL PROCEDURE Turkeys were fed the same diet in electrically heated battery brooders on raised
1956
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ABSTRACT Three experiments were conducted with B. B. Bronze and B. B. White turkeys, four to nine weeks of age. Poults were fed 12.5, 25,37.5, 50 or 75 p.p.m. (or 0.00125, 0.0025, 0.00375, 0.0050, and 0.0075%) of nifursol and given graded levels of exposure to histomoniasis. The exposure levels consisted of graded doses of infective material (embryonated Heterakis gallinae ova) administered orally. Histomoniasis mortality was recorded during post-exposure periods of 27-35 days; dead birds were examined for hepatic and cecal lesions typical for histomoniasis. Birds which survived the post-exposure period were sacrificed and examined for hepatic and cecal lesions. Graded doses of infective material caused graded levels of mortality and morbidity only in poults fed the unmedicated diet in two experiments. Dietary levels of 37.5, 50, 75 and 100 p.p.m. of nifursol were equally effective in preventing histomoniasis mortality and morbidity. In one experiment, both 12.5 and 25 p.p.m. of nifursol completely prevented mortality and morbidity among poults given graded doses of infective material. However, 20.8 percent mortality and 27.1 percent morbidity occurred among poults fed 12.5 p.p.m. of nifursol in another experiment. The minimum effective level of nifursol in feed for prevention of histomoniasis mortality and morbidity was between 25 and 50 p.p.m. Level of exposure or dose of infective material given poults had very little influence on the efficacy of nifursol treatments.
NlFURSOL AND HlSTOMONIASIS
on a serial dilution. Poults were assigned to dietary treatments at 30 days of age and given infective material five days later. Experiment 2. This experiment involved nifursol levels of 0, 12.5, 25 and 50 p.p.m.; infective doses were approximately 0, 20, 50 and 200 embryonated H. gallinae ova per bird based on a serial dilution. Poults were assigned to dietary treatments at 27 days of age and given infective material four days later. Experiment 3. Dietary levels of nifursol evaluated in this experiment were 0, 12.5, 37.5 and 75 p.p.m. These treatments were initiated when poults were 33 days of age. Birds receiving each dietary treatment were given the following doses of infective material: 0, 40, 80 and 800 embryonated H. gallinae ova per bird based on a serial dilution. Doses of infective material were divided in half and given when poults were 36 and 39 days of age. The split or divided dose of infective material was believed to provide a more uniform exposure within each dosage rate. RESULTS AND DISCUSSION
Experiment 1. Data from this experiment are presented in Table 1. Graded doses of infective material caused graded levels of mortality and morbidity in poults fed the unmedicated control diet (0 p.p.m. of nifursol). No mortality or morbidity occurred in poults receiving any of the nifursol treatments. Therefore, the lowest level of dietary nifursol was completely effective in preventing histomoniasis mortality and morbidity in this experiment. These results are in close agreement with data previously reported by Vatne et al. (1967, 1969). These workers reported that 0.005 percent of nifursol (50 p.p.m.) prevented all mortality in two experiments and allowed only 5.3 percent mortality in a third. It should be mentioned, however, that Vatne et al. (1969) used stronger exposures to histo-
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wire floors until they were 27 to 33 days of age. Both Broad Breasted (B.B.) Bronze and B. B. White poults were used. Six female and six male poults were randomly assigned to each treatment in all three experiments. A four by four factorial arrangement of treatments was employed in each experiment; all combinations of four dietary levels of nifursol and four doses of infective material were included each time. Nifursol levels were added to a 28 percent protein, practical-type, corn-soybean meal diet. Composition of this diet was reported by Bowen and Sullivan (1971). After receiving dietary treatments for three to five days, poults were exposed with embryonated Heterakis gallinae ova containing Histomonas meleagridis. Exposure was accomplished by placing the H. gallinae ova directly into the crop with a pipette. Postexposure experimental periods were 27, 28 and 35 days, respectively, in experiments 1, 2 and 3. All birds which died following exposure were examined for hepatic and cecal lesions typical for histomoniasis. All surviving poults were necropsied at the end of post-exposure periods and likewise examined for lesions. Mortality as reported herein includes birds which died with lesions typical for histomoniasis. Morbidity includes all birds (dead birds during the post-exposure period and survivors), which showed hepatic and/or cecal lesions typical for the disease. All mortality and morbidity data were subjected to analysis of variance following an arsin transformation of percentage data (Steel and Torrie, 1960). A Fisher's L.S.D. value was computed for main effect means relative to mortality and morbidity data in each experiment. Experiment 1. Nifursol levels evaluated in this experiment were 0, 50, 75 and 100 parts per million (p.p.m.); infective doses were approximately 0, 20, 50 and 100 embryonated H. gallinae ova per bird based
1957
1958
T. W. SULLIVAN, R. J. MITCHELL AND O. D. GRACE TABLE 1.—Mortality and morbidity data
TABLE 2.—Mortality and morbidity data
from experiment 1*
from experiment 2*
Nifursol in feed, p.p.m.
Nifursol in feed, p.p.m.
Avg. 50 B. gallinae ova/bird 0 20 50 100
Average
Average
0.0
% Mortality!
0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0
0.0a
0.0a
0.0a
% Morbidity1 0.0 0.0 0.0 0.0
0.0
25.0 75.0 83.0
0.0 0.0 0.0 0.0
45.8b
0.0a
0.0a
0.0a
o.o 0.0 0.0
0.0a 6.2b 18.8c 20.8c
0.0a 6.2b 18.8c 20.8c
* Six female and six male B. B. Bronze poults were randomly assigned to each of the 16 treatments at 30 days of age, and exposed to infective material at 35 days of age; surviving poults were weighed, sacrificed and examined at 62 days of age. 1 Composite means for nifursol levels or infective doses, which are followed by the same letter, within either mortality or morbidity data do not differ significantly (P <0.05) based on Fisher's L.S.D. test.
moniasis than any dose of infective material provided in this experiment of our study. Further experiments in this study were designed with lower levels of dietary nifursol and larger doses of infective material. Experiment 2. Lower levels of dietary nifursol, 12.5 and 25 p.p.m., completely prevented mortality, and with one exception, totally prevented morbidity in this trial (Table 2). One bird receiving 25 p.p.m. of nifursol showed hepatic and cecal lesions typical for histomoniasis. This bird survived the 28 day post-exposures period and was sacrificed; mild lesions were then detected during necropsy. The lowest level of nifursol in this experiment, 12.5 p.p.m., was more effective than reported by Vatne et al. (1969). Only 44.5 percent of poults receiving 12.5 p.p.m. of nifursol survived in an experiment conducted by Vatne and coworkers. The lower efficacy of 12.5 p.p.m. of nifursol in their study was most likely due to a stronger exposure (larger dose of infective material) employed as compared to the infective doses used in this experiment. Graded doses of infective material did result in graded levels of mortality and
B. gallinae ova/bird
12.5
25
% Mortality'
50
33.3 75.0 91.7
0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0
Average
50.0b
0.0a
0.0a
0.0a
0 20 50 200
33.3 75.0 91.7
0.0 0.0 0.0 0.0
0.0 0.0 0.0 8.3
0.0 0.0 0.0 0.0
50.0b
0.0a
2.1a
0.0a
0 20 50 200
Average
0.0
0.0
% Morbidity'
0.0a 8.3a 18.8b 22.9b
0.0a 8.3ab 18.8ac 25.0c
* Six female and six male B. B. White poults were randomly assigned to each of the 16 treatments at 27 days of age, and exposed to infective material at 31 days of age; surviving poults were weighed, sacrificed and examined at 59 days of age. 1 Composite means for nifursol levels or infective doses, which are followed by the same letter, within either mortality or morbidity data do not differ significantly (P<0.05) based on Fisher's L.S.D. test.
morbidity among poults receiving the unmedicated diet in this experiment. Experiment 3. Data from this experiment are presented in Table 3. Mortality among poults receiving 12.5 p.p.m., of dietary nifursol was 20.8 percent, when all infective doses were included; likewise, the incidence of morbidity was 27.1 percent among birds receiving this treatment. In contrast, the same level of nifursol completely prevented histomoniasis mortality and morbidity in experiment 2. Larger doses of infective material were employed in this experiment than in experiment 2; also, the culture of infective material used in this experiment could have been more potent. The two higher levels of nifursol, 37.5 and 75 p:p.m. provided almost complete protection against histomoniasis mortality. Only one bird within each of these treatments died during the experiment. Morbidity was slightly higher among birds receiving 37.5 p.p.m. of nifursol than in those receiving 75 p.p.m., this difference was not statistically significant. Graded doses of infective material, approximately 40, 80 and 800 H.
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0 20 50 100
0.0
25.0 75.0 83.0 45.8b
0
75
NlFURSOL AND HlSTOMONIASIS TABLE 3.—Mortality and morbidity data from experiment 3*
1959
to efficacy against histomoniasis in this experiment.
Nifursol in feed, p.p.m. • 0
U, gallinae ova/bird 0 40 80 800
Average
12.5
37.5
% Mortality1
0.0
0.0
83.3 83.3 100.0
33.3 25.0 25.0
0.0 0.0 8.3 0.0
0.0 0.0 8.3 0.0
20.8b
2.1a
2.1a
66.6c
Avg.
0.0a 29.2b 31.2b 31.2b
% Morbidityi 0 40 80 800
0.0
50.0 25.0 33.3
66.6c
27.1b
0.0 8.3 0.0
0.0 0.0 8.3 0.0
6.2a
2.1a
16.7
0.0a 35.4b 33.3b 33.3b
* Six female and six male B. B. White poults were randomly assigned to each of the 16 treatments at 33 days of age and exposed to infective material at 36 and 39 days of age; surviving poults were weighed, sacrificed and examined at 71 days of age. 1 Composite means for nifursol levels or infective doses, which are followed by the same letter, within either mortality or morbidity data do not differ significantly (P<0.05) based on Fisher's L.S.D. test.
nae ova/bird, did not produce graded levels of mortality and morbidity. The lowest infective dose caused 83.3 percent mortality in poults fed the unmedicated diet. There was no significant difference in either mortality or morbidity between poults fed 37.S and 75 p.p.m. of nifursol. These nifursol levels were statistically equivalent relative
This study was supported in part by a grant from Salsbury Laboratories, Charles City, Iowa. Nifursol is the generic name assigned to 3, 5-dinitrosalicylic acid, 5-nitrofurylidene hydrazide, a new drug under development by Salsbury Laboratories. REFERENCES Bowen, T. E., and T. W. Sullivan, 1971. Influence of dietary cupric sulfate on the response of young turkeys to penicillin-streptomycin (1:3). Poultry Sci. 50:273-278. Steel, R. G., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., New York. Sullivan, T. W., and O. D. Grace, 1968. Prophylactic efficacy of nifursol against histomoniasis in turkeys 4-9, 8-13 and 13-18 weeks of age. Poultry Sci. 47 : 1724. Vatne, R. D., R. R. Baron and N. F. Morehouse, 1967. Salfuride, a new antihistomonal compound. Poultry Sci. 46:1332. Vatne, R. D., R. R. Baron and N. F. Morehouse, 1969. Histomonastatic activity of nifursol in turkeys. Poultry Sci. 48: 590-596.
NEWS AND NOTES {Continued pom page 1955) Private Collections and Collectors Agway, Inc., Box 1333, Syracuse, New York 13201 Louis C. Arrington, 3821 St. Clair St., Madison, Wisconsin 53711 Sam Bejach, Gardner Hotel, P.O. Box 48, El Paso, Texas 79940 Eugene S. Carlson, 619 E. Chestnut St., Hoopeston, Illinois 60942 Donald C. Chandler, New Gloucester, Maine 04620 (Wyandottes only) Lloyd R. Champion, Poultry Science Department, Michigan State University, East Lansing, Michigan 48823 Lowndes W. Clark, 717 Ritchie Ave., Silver Spring, Maryland 20910
A A B C
B A
Cobb Breeding Corporation, Concord, Massachusetts 01742 Colonial Poultry Farms, Pleasant Hill, Missouri 64080 Everet T. Conway, 621 Water St., P.O. Box 449, Seaford, Delaware 19973 (Pictures and momentos of early Delmarva processing) DeKalb AgResearch, Sycamore Road, DeKalb, Illinois 60115 Roger Evenson, Box 5, Stanton, Minnesota 55081 John M. Freeman, 1808 Chestnut St., Philadelphia, Pennsylvania 19103 D. W. Francis, Poultry Science Department, New Mexico State University, Las Cruces, New Mexico 88001
(Continued on page 1962)
A C
A C A
A
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Average
0.0
83.3 83.3 100.0
ACKNOWLEDGMENT
75