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The Diagnostic Value of the Various Tests for Pullorum Disease
T h e Diagnostic Value of the Various Tests for Pullorum Disease W. A. H I G G I N S AND CARL H.
SCHROEDER*
Detroit, Michigan (Presented at Annual Meeting August 2-4, 1933)
ORKING independently of each other, two agencies in 1931 simultaneously reported, on the very promising results which they obtained from the rapid whole blood test for S. pullorum when using a stained antigen. Coburn and Stafseth (1931) developed a stained antigen after the method of Huddleson and Abell for the rapid serum agglutination test for abortion in cattle. Schaffer, MacDonald, Hall, and Bunyea (1931) also announced the results with a similar, stained antigen developed by these investigators of the Bureau of Animal Industry, U.S.D.A. The method of preparing the stained antigen has been patented by the U.S.D.A. and permits to produce it commercially have been granted to manufacturers meeting the requirements stipulated by the Bureau of Animal Industry. It is, of course, important to know whether the stained antigens prepared by the various licensed laboratories are equally satisfactory. To obtain first-hand information on this point was one of the objectives of this test. PLAN OF THE EXPERIMENT
The long, tube method was used as the standard for comparison. The antigen was supplied by Drs. D. R. Coburn and H. J. Stafseth of the Michigan State College. The following stained antigens were used by the loop method in conducting the rapid, whole-blood test: Antigen A: Coburn-Stafseth type stained * Larro Research Farm of General Mills, Inc.
antigen; furnished by the Michigan State College. Antigen B, C, D: Commercially available stained antigens, manufactured under federal license according to the procedure outlined by the Bureau of Animal Industry. Antigen E: Supplied by Dr. Bunyea of U.S.D.A. and prepared according to the method developed and patented by the Department. Such quantities of the antigen as were left over from the first test, were properly stored in an electric refrigerator at a constant temperature of 38°F., and were used in a repetition of the test four months later. The tests with the five stained antigens were made at the same time when the blood samples were taken for the tube test. The first test was made on January 10, 1933, on 108 White Leghorn pullets, nine months of age'. The pullets were from a brood of chicks purchased from a nearby flock. Although the early death rate in this brood was low, bacteriological examination of each dead chick demonstrated that pullorum infection was primarily responsible for the recorded losses. The following symbols were used in recording the readings: — = Negative ± = Doubtful + = Slight reaction + + = Strong reaction + + + = Very strong reaction, and in the case of the rapid test, also very quick reaction The accompanying table represents a
[239]
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii - Manoa on June 18, 2015
W
240
POULTRY
SCIENCE
TABLE 1.—Summarized comparisons 1st Test Tube Method
93 B i r d s -
A
B
C
D
E
-
89
67
91
93
89
80
86
52
90
89
79
+
4
6
2
3
9
4
25
2
1
6
2
1
13
3
5
1
2
2
+ ++ +++
19 1
1
6
3
+
1
+ ++ +++
2
-
4
+
1
+ ++ +++ +
A
B
C
D
E
1
1
6
5
5
5
5
1
1
1
5
2
4
4
1
2 1
1 6
5
6
3
5
5
3
1
1
2
5
5
5
1
1
1
1
1
1
1
2
2
2
2
2
2
1
1 1
2 Birds+ +
Rapid whole blood test with Stained Antigens:
1 2
+ ++ +++
2
1
1
1
1
1 1
1 1 1
1
+ 2 Birds+ + +
+ ++ +++
1 1
2
1
1
1
1
1
1
1
1
2
summary of the results obtained from the tween the results obtained by the tube test tube method as compared with each of the and the rapid, whole blood test with all but five tests with the stained antigen. one of the five stained antigens involved in the comparison. DISCUSSION OF DATA Of the commercial antigens, the two desFirst test.—The data presented demon- ignated C and D, respectively, proved as strate a very satisfactory agreement be- satisfactory as did the tube method or either
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii - Manoa on June 18, 2015
5 Birds+
Rapid whole blood test with Stained Antigens:
Tube Method
Reaction
6 Birds +
2nd Test (four months later)
JULY,
1934.
VOL. XIII,
No.
241 birds which reacted negatively or doubtfully might have been in the early stage of infection and, for this reason, failed to react to the agglutination test at that time. SUMMARY
Very close, but not quite complete, agreement was attained between the results from the tube agglutination test as compared with the rapid, whole-blood test with four of the five stained antigens in this test. Excepting the one lot of stained antigen which produced results at marked variance, the rapid, whole blood test with stained antigen was of equal diagnostic value as the tube agglutination test for 5. pullorum. ACKNOWLEDGMENT
The authors wish to thank Drs. D. R. Coburn and H. J. Stafseth as well as Dr. H. A. Bunyea for the co-operation extended by furnishing some of the antigens. REFERENCES Coburn, D. R. and H. J. Stafseth, 1931. A field test for pullorum disease. Jl. A.V.M.A.-N.S., 32:241-243. Schaffer, J. M., A. D. MacDonald, W. J. Hall, and H. Bunyea, 1931. A stained antigen for the rapid whole blood test for pullorum disease. Jl. A.V.M.A.-N.S., 32 :236-240
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii - Manoa on June 18, 2015
one of the two "official" antigens used. The agreement is more complete among the positive reactors and, therefore, a positive reaction may be considered a highly accurate indication of S. pullorum infection. The possibility exists that antigen B was not representative of the product generally put out by its manufacturer or that its efficiency became impaired through conditions beyond the maker's control. Second test.—When the test was repeated four months later on the same birds with the same antigens, again close, but not perfect, agreement was demonstrated among all antigens save "B." As in the first test, the latter again manifested marked discrepancies. A few of the birds which gave a negative or doubtful reaction in the first test, became doubtful or positive reactors in the second test. This shift does not reflect upon the stability of the antigens. On several occasions, the authors previously observed an increase in the incidence of suspicious and positive reactors in flocks from which the reactors were not removed. This is particularly true when males were left in the pens, as happened to be the case in this test. Furthermore, at the time of the first test some
4
SCHROEDER*
Detroit, Michigan (Presented at Annual Meeting August 2-4, 1933)
ORKING independently of each other, two agencies in 1931 simultaneously reported, on the very promising results which they obtained from the rapid whole blood test for S. pullorum when using a stained antigen. Coburn and Stafseth (1931) developed a stained antigen after the method of Huddleson and Abell for the rapid serum agglutination test for abortion in cattle. Schaffer, MacDonald, Hall, and Bunyea (1931) also announced the results with a similar, stained antigen developed by these investigators of the Bureau of Animal Industry, U.S.D.A. The method of preparing the stained antigen has been patented by the U.S.D.A. and permits to produce it commercially have been granted to manufacturers meeting the requirements stipulated by the Bureau of Animal Industry. It is, of course, important to know whether the stained antigens prepared by the various licensed laboratories are equally satisfactory. To obtain first-hand information on this point was one of the objectives of this test. PLAN OF THE EXPERIMENT
The long, tube method was used as the standard for comparison. The antigen was supplied by Drs. D. R. Coburn and H. J. Stafseth of the Michigan State College. The following stained antigens were used by the loop method in conducting the rapid, whole-blood test: Antigen A: Coburn-Stafseth type stained * Larro Research Farm of General Mills, Inc.
antigen; furnished by the Michigan State College. Antigen B, C, D: Commercially available stained antigens, manufactured under federal license according to the procedure outlined by the Bureau of Animal Industry. Antigen E: Supplied by Dr. Bunyea of U.S.D.A. and prepared according to the method developed and patented by the Department. Such quantities of the antigen as were left over from the first test, were properly stored in an electric refrigerator at a constant temperature of 38°F., and were used in a repetition of the test four months later. The tests with the five stained antigens were made at the same time when the blood samples were taken for the tube test. The first test was made on January 10, 1933, on 108 White Leghorn pullets, nine months of age'. The pullets were from a brood of chicks purchased from a nearby flock. Although the early death rate in this brood was low, bacteriological examination of each dead chick demonstrated that pullorum infection was primarily responsible for the recorded losses. The following symbols were used in recording the readings: — = Negative ± = Doubtful + = Slight reaction + + = Strong reaction + + + = Very strong reaction, and in the case of the rapid test, also very quick reaction The accompanying table represents a
[239]
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii - Manoa on June 18, 2015
W
240
POULTRY
SCIENCE
TABLE 1.—Summarized comparisons 1st Test Tube Method
93 B i r d s -
A
B
C
D
E
-
89
67
91
93
89
80
86
52
90
89
79
+
4
6
2
3
9
4
25
2
1
6
2
1
13
3
5
1
2
2
+ ++ +++
19 1
1
6
3
+
1
+ ++ +++
2
-
4
+
1
+ ++ +++ +
A
B
C
D
E
1
1
6
5
5
5
5
1
1
1
5
2
4
4
1
2 1
1 6
5
6
3
5
5
3
1
1
2
5
5
5
1
1
1
1
1
1
1
2
2
2
2
2
2
1
1 1
2 Birds+ +
Rapid whole blood test with Stained Antigens:
1 2
+ ++ +++
2
1
1
1
1
1 1
1 1 1
1
+ 2 Birds+ + +
+ ++ +++
1 1
2
1
1
1
1
1
1
1
1
2
summary of the results obtained from the tween the results obtained by the tube test tube method as compared with each of the and the rapid, whole blood test with all but five tests with the stained antigen. one of the five stained antigens involved in the comparison. DISCUSSION OF DATA Of the commercial antigens, the two desFirst test.—The data presented demon- ignated C and D, respectively, proved as strate a very satisfactory agreement be- satisfactory as did the tube method or either
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii - Manoa on June 18, 2015
5 Birds+
Rapid whole blood test with Stained Antigens:
Tube Method
Reaction
6 Birds +
2nd Test (four months later)
JULY,
1934.
VOL. XIII,
No.
241 birds which reacted negatively or doubtfully might have been in the early stage of infection and, for this reason, failed to react to the agglutination test at that time. SUMMARY
Very close, but not quite complete, agreement was attained between the results from the tube agglutination test as compared with the rapid, whole-blood test with four of the five stained antigens in this test. Excepting the one lot of stained antigen which produced results at marked variance, the rapid, whole blood test with stained antigen was of equal diagnostic value as the tube agglutination test for 5. pullorum. ACKNOWLEDGMENT
The authors wish to thank Drs. D. R. Coburn and H. J. Stafseth as well as Dr. H. A. Bunyea for the co-operation extended by furnishing some of the antigens. REFERENCES Coburn, D. R. and H. J. Stafseth, 1931. A field test for pullorum disease. Jl. A.V.M.A.-N.S., 32:241-243. Schaffer, J. M., A. D. MacDonald, W. J. Hall, and H. Bunyea, 1931. A stained antigen for the rapid whole blood test for pullorum disease. Jl. A.V.M.A.-N.S., 32 :236-240
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii - Manoa on June 18, 2015
one of the two "official" antigens used. The agreement is more complete among the positive reactors and, therefore, a positive reaction may be considered a highly accurate indication of S. pullorum infection. The possibility exists that antigen B was not representative of the product generally put out by its manufacturer or that its efficiency became impaired through conditions beyond the maker's control. Second test.—When the test was repeated four months later on the same birds with the same antigens, again close, but not perfect, agreement was demonstrated among all antigens save "B." As in the first test, the latter again manifested marked discrepancies. A few of the birds which gave a negative or doubtful reaction in the first test, became doubtful or positive reactors in the second test. This shift does not reflect upon the stability of the antigens. On several occasions, the authors previously observed an increase in the incidence of suspicious and positive reactors in flocks from which the reactors were not removed. This is particularly true when males were left in the pens, as happened to be the case in this test. Furthermore, at the time of the first test some
4