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Agglutinin Content of Eggs from Salmonella Pullorum Infected Hens
Agglutinin Content of Eggs from Salmonella Pullorum Infected Hens NORMA A. FRANK AND B. H. EDGINGTON
Ohio Agricultural Experiment Station, Division of Animal Industry, Animal Disease Laboratories, Reynoldsburg, Ohio (Received for publication June 22, 1937)
B
EAUDETTE (1), in 1923, demonstrated the presence of specific agglutinins in the albumin of eggs from B. pullorum infected hens. Agglutination was most marked in 1:8 dilution of the egg albumin but occurred in some cases in a dilution of 1:64. No correlation of the blood serum and egg albumin titers was reported. May (3), in 1924, using hens inoculated with cultures of S. pullorum, P. avicida and S. gallinarum found specific agglutinins in the blood serum of all birds except those inoculated with P. avicida. The agglutinin titer of albumin from eggs laid by hens inoculated with 5. pullorum and S. gallinarum was lower than their respective blood titers. Paille (4), in 1935, reported that the agglutinin titer of yolk of eggs from the hens infected with fowl typhoid was practically the same as that of their blood serum or the whole blood. Agglutinins were not demonstrated in the whites of these eggs. Jansen (2), in 1936, reported that blood of hens infected with 5. enteritidis var. essen and 5. typhi-murium, which gave a strongly positive reaction, showed similar reactions with yolk material, but when the whole blood gave a weak reaction, negative results were obtained with the yolk. He also reports the presence of agglutinins in the yolks of eggs from hens infected with S. pullorum.
Studies were undertaken to determine the constancy of agglutinins in eggs produced by S. pullorum infected hens. The stained-antigen and technic of the whole-blood test as recommended by Schaeffer, MacDonald, Hall, and Bunyea (5) were used. A standard loop of egg yolk or albumin was substituted for that of whole blood. TEST NO. 1
Two 18-month-old White Rock pullorumdisease reacting hens were used in this test. Each bird gave a positive reaction to the whole-blood test. The agglutinin titer as demonstrated by the tube test was partial at 1:800 for one hen and partial at 1:400 for the other. These birds were not trapnested, but were under observation over a period of about five months, during which time 32 eggs were layed and tested. Positive reactions were obtained with the yolks and negative reactions with the albumin of each egg. Discolored ova obtained from these hens on autopsy were cultured for 5. pullorum; the apparently normal ova were tested for S. pullorum agglutinins. These organisms and agglutinins were demonstrated in material from each hen. TEST NO. 2 Nineteen hens positive to the whole-blood test were tested by the tube method and showed titers ranging from incomplete at 1:50 to partial at 1:1600. These birds were
[442]
NOVEMBER,
1937.
VOL.
XVI,
trapnested over a period of seven and onehalf months. A total of 463 eggs were tested. The results of these tests are given in Table 1.
No.
443
6
The hens were autopsied at the termination of the experiment and any discolored ova observed were cultured. Isolated organisms which culturally and morphologically
TABLE 1.—Agglutination reactions of whites and yolks of eggs from pullorum-reactor hens Agglutination of blood Hen no.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Tube test at Whole blood beginning of test at time of autopsy experiment 50 100 200 200 800 200 800 200 1600 400 100 100 100 50 400 800 200 1600 400
P* P I C P I C I P C P P P P I I I I P
C C C C C C C C C C C C C C C C C C C
Agglutination of yolks Total eggs tested 32 15 23 9 50 15 17 47 45 27 22 8 6 23 29 20 16 34 25
C
P
I
T
7 11 10 3 49 7 11 41 45 27 16 3 6 9 29 20 16 31
14 1 12 2 1 7 5 5
7 3 1 3
4
N
1
1 1 1
6 5 4
7
3
1
23
2 1
1
Recovery of salmonella pullorum from ova
Agglutination of immature ova
+ + + + + + + + + + + + + + + + + +
C C P C C C C C C C C P C P C C C C C
* C—Complete agglutination; I—Incomplete=75 percent agglutination; P—Partial=50 percent agglutination; T—Trace=25 percent agglutination; N—No agglutination.
The yolks of 341 eggs gave complete, 63 incomplete, 49 partial, and 10 a trace of agglutination. Agglutination was not observed with the albumin of any of these eggs. The clumps of organisms were larger in rapidly agglutinating reactions than in those agglutinating slowly. Usually when the yolk agglutination occurred rapidly with comparatively large agglutination particles the blood titer of the respective bird was high. One exception was noted; none of the 25 eggs layed by a Barred Rock hen whose blood titer was incomplete at 1:400, gave complete reactions. Yolks of 92 eggs layed by these hens were cultured for S. pullorum prior to their use in the agglutination test. All media inoculated with this yolk material remained sterile.
resembled S. pullorum were used as antigen for agglutination tests with known positive and negative pullorum sera. Judged by this criterion 5. pullorum was recovered from 18 of the 19 hens. The hen from which S. pullorum was not recovered had a blood titer incomplete at 1:50. In tests of 23 eggs which she produced, nine gave complete, four incomplete, and seven partial agglutinations while three showed only a trace of agglutinations. Yolk material, collected at the time of autopsy of the hens, from apparently normal but incompletely developed ova gave positive agglutination reactions in 16 hens and incomplete in three. One of the latter was the hen from which S.. pullorum was not isolated. SUMMARY Yolk and albumin from 495 eggs layed
444 b y 21 h e n s h a v i n g S. pullorum
P O U L T R Y
t i t e r s of their
blood serum ranging from incomplete at 1:50 to partial at 1:1600 were tested by the stained-antigen-plate method. Agglutinins were demonstrated in the yolk of each egg but in no instance in the albumin. Complete agglutination of the antigen occurred regularly with yolk material from eggs produced by six of these hens. Eggs from the other hens gave reactions that varied from complete to a mere trace of agglutination. Ninety-two of these eggs, the total number layed during October, November, and December, were cultured for S. pullorum with negative results. Blood collected from each of the 21 hens at the time of autopsy gave complete agglutinations in the whole-blood-plate tests. S. pullorum cultures were obtained from discolored ova of 20 of the 21 hens. Material obtained at the time of autopsy from apparently normal but incompletely
S C I E N C E
developed ova gave complete agglutination with yolk from 18 of the hens and incomplete with that from three. Agglutination of the antigen usually was most rapid and the clumps largest with material from the high-titer birds. REFERENCES
1. Beaudette, F. R. 1923. Agglutinins for Bacterium Pullorum in Hen's Eggs. Jour. Immunology, 8:493-499. 2. Jansen, J. 1936. Agglutination of Yolk Substance with Salmonella of Ducks. Dtsch. tieraztl. Wschr. 19:340-341. 3. May, H. G. 1924. The Examination of Eggs from Infected and Immunized Hens with Germicidal Tests on Albumin and Blood Serum. Bull. 197. Agr. Expt. Sta. of Rhode Island State. 4. Paille, R. 193S. Avian Typhoid and the Ability of Eggs to Agglutinate. Rev. Vet. et Jour, de Med. Vet., 87 :S78. 5. Schaeffer, J. M., A. D. MacDonald, W. J. Hall, H. Bunyea, 1931. A Stained Antigen for the Rapid Whole Blood Test for Pullorum Disease. Jour. Am. Vet. Med. Ass., 79:236-240.
Ohio Agricultural Experiment Station, Division of Animal Industry, Animal Disease Laboratories, Reynoldsburg, Ohio (Received for publication June 22, 1937)
B
EAUDETTE (1), in 1923, demonstrated the presence of specific agglutinins in the albumin of eggs from B. pullorum infected hens. Agglutination was most marked in 1:8 dilution of the egg albumin but occurred in some cases in a dilution of 1:64. No correlation of the blood serum and egg albumin titers was reported. May (3), in 1924, using hens inoculated with cultures of S. pullorum, P. avicida and S. gallinarum found specific agglutinins in the blood serum of all birds except those inoculated with P. avicida. The agglutinin titer of albumin from eggs laid by hens inoculated with 5. pullorum and S. gallinarum was lower than their respective blood titers. Paille (4), in 1935, reported that the agglutinin titer of yolk of eggs from the hens infected with fowl typhoid was practically the same as that of their blood serum or the whole blood. Agglutinins were not demonstrated in the whites of these eggs. Jansen (2), in 1936, reported that blood of hens infected with 5. enteritidis var. essen and 5. typhi-murium, which gave a strongly positive reaction, showed similar reactions with yolk material, but when the whole blood gave a weak reaction, negative results were obtained with the yolk. He also reports the presence of agglutinins in the yolks of eggs from hens infected with S. pullorum.
Studies were undertaken to determine the constancy of agglutinins in eggs produced by S. pullorum infected hens. The stained-antigen and technic of the whole-blood test as recommended by Schaeffer, MacDonald, Hall, and Bunyea (5) were used. A standard loop of egg yolk or albumin was substituted for that of whole blood. TEST NO. 1
Two 18-month-old White Rock pullorumdisease reacting hens were used in this test. Each bird gave a positive reaction to the whole-blood test. The agglutinin titer as demonstrated by the tube test was partial at 1:800 for one hen and partial at 1:400 for the other. These birds were not trapnested, but were under observation over a period of about five months, during which time 32 eggs were layed and tested. Positive reactions were obtained with the yolks and negative reactions with the albumin of each egg. Discolored ova obtained from these hens on autopsy were cultured for 5. pullorum; the apparently normal ova were tested for S. pullorum agglutinins. These organisms and agglutinins were demonstrated in material from each hen. TEST NO. 2 Nineteen hens positive to the whole-blood test were tested by the tube method and showed titers ranging from incomplete at 1:50 to partial at 1:1600. These birds were
[442]
NOVEMBER,
1937.
VOL.
XVI,
trapnested over a period of seven and onehalf months. A total of 463 eggs were tested. The results of these tests are given in Table 1.
No.
443
6
The hens were autopsied at the termination of the experiment and any discolored ova observed were cultured. Isolated organisms which culturally and morphologically
TABLE 1.—Agglutination reactions of whites and yolks of eggs from pullorum-reactor hens Agglutination of blood Hen no.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Tube test at Whole blood beginning of test at time of autopsy experiment 50 100 200 200 800 200 800 200 1600 400 100 100 100 50 400 800 200 1600 400
P* P I C P I C I P C P P P P I I I I P
C C C C C C C C C C C C C C C C C C C
Agglutination of yolks Total eggs tested 32 15 23 9 50 15 17 47 45 27 22 8 6 23 29 20 16 34 25
C
P
I
T
7 11 10 3 49 7 11 41 45 27 16 3 6 9 29 20 16 31
14 1 12 2 1 7 5 5
7 3 1 3
4
N
1
1 1 1
6 5 4
7
3
1
23
2 1
1
Recovery of salmonella pullorum from ova
Agglutination of immature ova
+ + + + + + + + + + + + + + + + + +
C C P C C C C C C C C P C P C C C C C
* C—Complete agglutination; I—Incomplete=75 percent agglutination; P—Partial=50 percent agglutination; T—Trace=25 percent agglutination; N—No agglutination.
The yolks of 341 eggs gave complete, 63 incomplete, 49 partial, and 10 a trace of agglutination. Agglutination was not observed with the albumin of any of these eggs. The clumps of organisms were larger in rapidly agglutinating reactions than in those agglutinating slowly. Usually when the yolk agglutination occurred rapidly with comparatively large agglutination particles the blood titer of the respective bird was high. One exception was noted; none of the 25 eggs layed by a Barred Rock hen whose blood titer was incomplete at 1:400, gave complete reactions. Yolks of 92 eggs layed by these hens were cultured for S. pullorum prior to their use in the agglutination test. All media inoculated with this yolk material remained sterile.
resembled S. pullorum were used as antigen for agglutination tests with known positive and negative pullorum sera. Judged by this criterion 5. pullorum was recovered from 18 of the 19 hens. The hen from which S. pullorum was not recovered had a blood titer incomplete at 1:50. In tests of 23 eggs which she produced, nine gave complete, four incomplete, and seven partial agglutinations while three showed only a trace of agglutinations. Yolk material, collected at the time of autopsy of the hens, from apparently normal but incompletely developed ova gave positive agglutination reactions in 16 hens and incomplete in three. One of the latter was the hen from which S.. pullorum was not isolated. SUMMARY Yolk and albumin from 495 eggs layed
444 b y 21 h e n s h a v i n g S. pullorum
P O U L T R Y
t i t e r s of their
blood serum ranging from incomplete at 1:50 to partial at 1:1600 were tested by the stained-antigen-plate method. Agglutinins were demonstrated in the yolk of each egg but in no instance in the albumin. Complete agglutination of the antigen occurred regularly with yolk material from eggs produced by six of these hens. Eggs from the other hens gave reactions that varied from complete to a mere trace of agglutination. Ninety-two of these eggs, the total number layed during October, November, and December, were cultured for S. pullorum with negative results. Blood collected from each of the 21 hens at the time of autopsy gave complete agglutinations in the whole-blood-plate tests. S. pullorum cultures were obtained from discolored ova of 20 of the 21 hens. Material obtained at the time of autopsy from apparently normal but incompletely
S C I E N C E
developed ova gave complete agglutination with yolk from 18 of the hens and incomplete with that from three. Agglutination of the antigen usually was most rapid and the clumps largest with material from the high-titer birds. REFERENCES
1. Beaudette, F. R. 1923. Agglutinins for Bacterium Pullorum in Hen's Eggs. Jour. Immunology, 8:493-499. 2. Jansen, J. 1936. Agglutination of Yolk Substance with Salmonella of Ducks. Dtsch. tieraztl. Wschr. 19:340-341. 3. May, H. G. 1924. The Examination of Eggs from Infected and Immunized Hens with Germicidal Tests on Albumin and Blood Serum. Bull. 197. Agr. Expt. Sta. of Rhode Island State. 4. Paille, R. 193S. Avian Typhoid and the Ability of Eggs to Agglutinate. Rev. Vet. et Jour, de Med. Vet., 87 :S78. 5. Schaeffer, J. M., A. D. MacDonald, W. J. Hall, H. Bunyea, 1931. A Stained Antigen for the Rapid Whole Blood Test for Pullorum Disease. Jour. Am. Vet. Med. Ass., 79:236-240.