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Mycoplasma synoviae and Mycoplasma gallisepticum Infections: Differentiation by the Hemagglutination-Inhibition Test
I N J E C T I O N OX V I T A M I N S AND N E O M Y C I N
and treated. Holding birds for three days significantly reduced nine-day growth to a b o u t 9 0 % of t h a t of other holding treatments. This difference is probably due to the total number of days on feed, which indicates t h a t the earlier feeding h a d been most beneficial by adding extra days on feed. Mortality was slightly b u t not consistently highest in the control group.
157
tribute to early poult mortality in control, untreated groups. ACKNOWLEDGMENT T h e supply of water-soluble vitamins and neomycin was supplied through the courtesy of Nopco Chemical Company. Appreciation is also extended to Hoffmann-La Roche for their generous supply of vitamins for the basal diet.
SUMMARY REFERENCES Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Harper, J. A., and W. E. Babcock, 1953. The effect of penicillin on early mortality and growth in poults. Poultry Sci. 32: 179-180. Jensen, L. S., J. Allred, R. Fry and J. McGinnis, 1956. Effect of vitamin injections on survival of chicks with a high mortality syndrome. Fed. Proc. 15: 558. National Research Council, 1966. Nutrient Requirements of Domestic Animals. I. Nutrient Requirements of Poultry. National Academy of Science, Washington, D. C. Nestor, K. E., and K. I . Brown, 1966. Holding poults after hatching reduces early growth rate. Ohio Agri. Expt. Report 51: 36. Snedecor, G. W., and W. G. Cochran, 1967. Statistical Methods. Sixth edition. Iowa State University Press, Ames, Iowa. 593 pp.
Mycoplasma synoviae and Mycoplasma gallisepticum Infections: Differentiation by the HemagglutinationInhibition Test T. H. VARDAMAN 1 AND HARRY W. YODER, JR.2 United States Department of Agriculture, Animal Disease and Parasite Research Division (Received for publication August 18, 1969)
O
N E agent responsible for the clinical syndrome in chickens known as infectious synovitis, cultivated b y Chalquest and Fabricant (1960), was designated 1 South Central Poultry Research Laboratory, State College, Mississippi 39762. 2 Southeast Poultry Research Laboratory, Athens, Georgia 30601.
Mycoplasma synoviae by Olson et al. (1964). An important criterion for determining whether M. synoviae infection is present in a flock is the presence or absence of specific antibody as demonstrated by serologic examination. A M. synoviae serum plate agglutination test antigen has been produced (Olson et al.,
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Turkey poults were pre-fed, when hatched or u p to three days after hatching, utilizing a technique of oral-forced pre-feeding of solutions as follows: plain tap water, vitamin, or vitamin plus neomycin. Then poults were given conventional diets and management, and monitored for 7 to 11 days. Significant increases in body weight usually resulted from pre-feeding the vitamin mixture or the vitamin plus neomycin solution as compared with no pre-feeding. A significant reduction in mortality during this period was recorded; mortality was usually lowest for those groups receiving the vitamin plus neomycin oral feeding. Dehydration appeared to substantially con-
158
T. H. VARDAMAN AND H. W. YODER, JR.
This study was done to determine if the M. synoviae and M. gallisepticum HI tests could be used to differentiate between M. synoviae and M. gallisepticum infections. MATERIALS AND METHODS
The chicken serums were obtained from birds infected by: 1) injecting isolate WVU 1853 or ISU isolate 1331 M. synoviae egg-yolk material into the hock joint or foot pad; 2) contact with birds infected with M. synoviae; 3) injecting M. gallisepticum egg-yolk into the airsacs; 4) contact with birds infected with M. gallisepticum; and 5) natural infection. The artificially infected chickens were grown in Horsfall and Bauer (1940) type isolation cabinets as modified by Drury et al. (1969). Serum samples from groups 1, 2, and 3 were from chickens inoculated at 3 weeks of age via foot pad or hock joint with 0.2 ml. of a 1:5, 1:25, or 1:100 dilution of M. synoviae egg-yolk. Serum
samples were obtained 5, 10, and 15 weeks postinoculation. Groups 4, 5, and 6 were serums from chickens that had been in contact with M. synoviae-mocvla.X.eA birds for 5, 10, and 15 weeks, respectively. Serum samples from group 9 were from turkeys that were inoculated at 5 weeks of age via air sac and sinus with 0.1 ml. of a 24-hour broth culture of M. synoviae strain 6130. Serum samples were obtained 5 weeks postinoculation. Serum samples from groups 11, 12, and 13 were from chickens inoculated at 3 weeks of age via air sacs with 0.2 ml. of M. gallisepticum undiluted egg-yolk. Serum samples were obtained 5, 8, and 11 weeks postinoculation. The methods of producing the M. synoviae hemagglutinating antigen and the procedure for the hemagglutinationinhibition test have been described (Vardaman and Yoder, 1969). The serums were tested by the HI test using M. synoviae and M. gallisepticum HA antigens, and the rapid serum plate test using M. synoviae serum plate test antigen and M. gallisepticum serum plate test antigen. The results of the HI and rapid serum plate agglutination tests were compared. RESULTS AND DISCUSSION
The serologic results from M. synoviaeand M. gallisepticum-miected birds are shown (Table 1). There were two different birds of the M. synoviae artificiallyinoculated group that had M. gallisepticum HI titers of 1:40 only. Bird 2012, from group 1, had an M. gallisepticum HI titer of 1:40 and a negative reaction to the M. gallisepticum serum plate test. It had an M. synoviae HI titer of 1:80 and a 3 + reaction to M. synoviae serum plate test. Bird 2077, from group 2, had an M. gallisepticum HI titer of 1:40 and a negative reaction to M. gallisepticum serum
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1963; and Roberts and Olesiuk, 1967). Olsen et al. (1964), and Roberts and Olesiuk (1967) reported that M. synoviaepositive serums would sometimes cause a serologic reaction with M. gallisepticum serum plate antigen. M. synoviae serum plate antigen is the only antigen commercially available for serologic examination for M. synoviae antibodies. Vardaman and Yoder (1969) reported the preparation of a M. .rywcwae-hemagglutinatmg antigen that showed good specificity when used in the hemagglutination-inhibition (HI) test. The hemagglutinationinhibition test is an official test, and is the most accurate serologically, for detecting antibodies against M. gallisepticum. It would seem desirable to have an HI test available for M. synoviae so that diagnostic laboratories could differentiate between M. synoviae and M. gallisepticum infections.
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SYNOVIAE
159
AND GALLISEPTICUM
TABLE 1.—Results of survey of serologic differentiation between M. synoviae and M. gallisepticum
Group
li
chicken chicken chicken chicken chicken chicken chicken chicken turkey turkey chicken chicken chicken chicken turkey
Type infec.
Ms. Ms. Ms. Ms. Ms. Ms. Ms. Ms.+Mg. Ms. Ms. Mg. Mg. Mg. Mg. Mg.
Meth. infec.
Art. Art. Art. Cont. Cont. Cont. Field Field Art. Field Art. Art. Art. Cont. Cont.
Total No. birds 111 99 33 19 14 20 30 24 16 19 46 42 24 70 3
H I titer of 1 :40 & above M. synoviae %
Mean
82.9 94.9 93.9 21.1 78.6 90.0 83.3 91.7 68.8 78.9 0.0 0.0 0.0 0.0 0.0
238.3 291.9 238.7 110.0 138.2 248.8 134.4 225.5 225.5 304.0 0.0 0.0 0.0 0.0 0.0
M. gallisepticum % 0.9 1.01 0.0 0.0 0.0 5.0 0.0 87.5 0.0 0.0 100.0 100.0 100.0 100.0 100.0
Serum plate agglutination test—positive M. synoviae
Mean
%
40.0 40.0 0.0 00.0 00.0 40.0 00.0 81.9 00.0 00.0 219.1 310.5 246.7 421.7 266.7
98.20 98.90 100.00 42.11 92.86 85.00 86.66 100.00 25.00 100.00 58.70 16.67 4.17 0.00 66.67
No. 109 98 33 8 13 17 26 24 4 13/135 27 7 1 0 2
M. gallisepticum
% 0.00 0.00 12.10 0.00 0.00 0.00 83.33 95.83 0.00 69.20 95.65 100.00 100.00 94.30 100.00
No. 0 0 4 0 0 0 25 23 0 9/135 44 42 24 66 3
i2 Groups 1, 2, and 3 were the same birds, with serum samples obtained 5, 10, and 15 weeks postinoculation with M. synoviae. Groups 4, 5, and 6 were the same birds, with serum samples obtained 5,10, and 15 weeks post contact with M. synoviae-inoculated birds. 3 Group 9 were turkeys inoculated via sinus and trachea at 5 weeks, with serum samples obtained 5 weeks postinoculation. 4 Groups 11,12, and 13 were the same birds, with serum samples obtained 5,8, and 11 weeks postinoculation with M. gallisepticum. 5 Number positive over the number tested.
plate test, but it had an M. synoviae HI titer of 1:640 and a 3 + reaction to M. synoviae serum plate test. Group 2 of the M. synoviae artificially-inoculated birds, from which samples were obtained at 10 weeks postinoculation, had a higher percentage of M. synoviae H I titers of 1:40 and a higher mean than groups 1 and 3 from which serum samples were obtained at 5 and 15 weeks postinoculation, respectively. The mean M. synoviae H I titers were 238.3,291.9, and 238.7 from groups 1, 2, and 3, respectively. The lower percentage of M. synoviae H I positive results and the mean M. synoviae HI titers of group 3, when compared to group 2, are due to the fact that most of the obviously sick birds either died or were killed before 15 weeks. Typical reactions of M. synoviae artificially-inoculated birds were two which had M. synoviae HI titers of 1:40, 1:80, and 1:320 at 5, 10, and 15 weeks postinoculation, respectively. Bird 1608, from group 6, had an M. gallisepticum HI titer of 1:40 and a negative reaction to the M. gallisepticum serum plate test, but it had an M. synoviae HI titer of 1:640 and a 4 + reaction to M.
synoviae serum plate test. The percentage of M. synoviae HI titers of 1:40 and above, and the mean HI titers of the M. synoviae contact birds increased with time as shown in Table 1, by comparing groups 4, 5, and 6. Group 7, field samples of M. synoviae suspects, did not show any reactions to M. gallisepticum HI test and had a mean M. synoviae H I titer of 134.4. There were many M. gallisepticum serum plate crossreactions since these samples were selected to confirm M. synoviae infection by using the HI test. Group 8 were field samples that had positive reactions to both the M. synoviae and M. gallisepticum HI test and serum plate tests. This flock had been diagnosed as having M. synoviae plate- and Hl-positive reactions and M. gallisepticum plateand Hi-negative reactions previously. Three months later, when the 24 serum samples were taken, this flock began to show respiratory signs. Serologic results at this time showed that the flock was infected with M. gallisepticum and M. synoviae. Group 9 were serum samples from tur-
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21 31 42 52 62 7 8 93 10 11« 12* 13« 14 15
Species
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T . H . VARDAMAN AND H . W. Y O D E R , J R .
keys t h a t had been inoculated with a broth culture of M. synoviae via sinus and trachea 5 weeks previously. Approximately 6 9 % of the birds had a M. synoviae H I titer of 1:40 and above with a mean titer of 225.5. Only 25.0% h a d a reaction to the M. synoviae serum plate test and they had 1 + reactions or very slight reactions. No turkey serums reacted to the M. gallisepticum H I test or serum plate test.
T h e d a t a show considerable cross reactions to M. synoviae and M. gallisepticum serum plate tests in field samples from chickens and turkeys. T h e y show a need for the M. synoviae H I test which, used in conjunction with the M. gallisepticum H I test, would aid the diagnostician in differentiating between M. synoviae and M. gallisepticum infections.
Group 10 were selected turkey serums from the field flocks sent to the laboratory for confirmation of results. They had strong M. synoviae serum plate reactions and h a d a mean M. synoviae H I titer of 304.0. The M. gallisepticum serum plate reactions were weak, a 1 + or 2 + , and all were negative to M. gallisepticum HI test. Groups 11, 12, and 13 were artificially infected with undiluted M. gallisepticum egg-yolk material b y injecting the material into the air sacs. T h e y were all positive to the M. gallisepticum H I test with a mean H I titer of 219.1, 310.5, and 246.7 at 5, 8, and 11 weeks postinoculation. There were no cross reactions to M. synoviae H I test. There were 58.7, 16.7, and 4 . 2 % reactors to the M, synoviae serum plate test compared to 95.6, 100.0, and 100.0% reactors to M. gallisepticum serum plate test, a t 5, 8, and 11 weeks postinoculation. The initial cross reactions to the M. synoviae serum plate test m a y be caused b y heavy yolk inoculation to infect the birds.
T h e hemagglutination-inhibition tests for Mycoplasma synoviae and M. gallisepticum were used on serum samples from artificially-infected, contact-exposed and normal chickens raised in isolation cabinets. Other samples from field flocks of chickens and turkeys were also tested. Antigens prepared for both M. synoviae a n d M. gallisepticum had good specificity in differentiating between these infections by the hemagglutination-inhibition procedure. Cross reactions detected by the rapid serum plate test procedure were not a problem in the H I test. T h e tests worked equally well with both chicken and turkey serums, and should be valuable for diagnostic purposes.
Group 14, M. gallisepticum contact chickens, reacted to M. gallisepticum H I and serum plate test only. Group 15, M. gallisepticum contact turkeys, show M. gallisepticum H I and serum plate positive reactions. They had negative results to M. synoviae H I test but some reacted to M. synoviae serum plate test.
Chalquest, R. R., and J. Fabricant, 1960. Pleuropneumonia-like organisms associated with synovitis in fowls. Avian Diseases, 4: 515-539. Drury, L. N., C. W. Beard and S. R. Hopkins, 1969. An isolation cabinet system for avian disease research. Avian Diseases, 13: 400-412. Horsfall, F. L., Jr., and J. H. Bauer, 1940. Individual isolation of infected animals in a single room. J. Bact. 40: 569-580. Olson, N. O., K. M. Kerr and A. Campbell, 1963. Control of infectious synovitis. 12. Preparation
SUMMARY
T h e authors express their appreciation to Mrs. Charmian Lee Gatlin a n d Mr. Boyd Turner, medical technologists, for their assistance. REFERENCES
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ACKNOWLEDGEMENTS
MYCOPLASMA SYNOVIAE AND GALLISEPTICUM of an agglutination test antigen. Avian Diseases, 7: 310-317. Olson, N. O., K. M. Kerr and A. Campbell, 1964. Control of infectious synovitis. 13. The antigen study of three strains. Avian Diseases, 8: 209214. Roberts, D. H., and 0. M. Olesiuk, 1967. Serological
161
studies with Mycoplasma synoviae. Avian Diseases, 11: 104-119. Vardaman, T. H., and H. W. Yoder, Jr., 1969. Preparation of Mycoplasma synoviae hemagglutinating antigen and its use in the hemagglutinationinhibition test. Avian Diseases, 13: 654-661.
ERIC L. MORGAN2 AND FRANK R. MRAZ Agricultural Research Laboratory of the University of Tennessee, Oak Ridge, Tennessee 37830 (Received for publication August 18, 1969)
V
ARIATIONS in time and level of maximum thyroidal uptake or biological half-life of 181I following a single dose have been reported in several species of birds (McFarland et al., 1966; Rosenberg et al., 1963a; Premachandra et al., 1958; Wahlberg, 1955). Featherston et al. (1966) found that chicks fed an iodinedeficient diet retained more 131I originally than did control chicks. Thyroids of iodine-deficient chicks trapped a maximum of 4 1 % of the original dose at 2 hr. with a biological half-life of 15 hr. Control chicks, however, trapped a maximum of only 4.5% at 8 hr with a biological half-life of more than 96 hr. The primary objectives of this study were to determine the effects of dietary iodine on the maximum uptake and biological half-life of 131I in the thyroid glands of Coturnix and Bobwhite quail and to 1 This manuscript is published with the permission of the Director of The University of Tennessee Agricultural Experiment Station, Knoxville, Tennessee. 2 Present Address: Department of Biology, The University of Tennessee, Chattanooga, Tennessee. 3 Operated by the Tennessee Agricultural Experiment Station for the U. S. Atomic Energy Commission under Contract No. AT-40-1-GEN-242.
compare the response of these two species with respect to thyroidal 13lI metabolism. EXPERIMENTAL All quail used in these studies were hatched from breeding stocks at the UTAEC Agricultural Research Laboratory. The Coturnix were originally acquired from Auburn University and the Bobwhites from the Tennessee Wildlife Commission's Buffalo Springs Game Bird Farm. Day-old Coturnix and Bobwhite chicks were placed in specially constructed 1/4-inch wire mesh screen cages adapted to jfelectrically-heated commercial chick brooders. They were immediately fed the experimental diets and at about 2 weeks of age moved to standard chick brooders. Half the birds in each experiment were fed the pheasant and quail starter diet of Scott et al. (1960) minus the Ca(IOs)2. The remaining half were fed this diet supplemented with 0.1 mg. iodide from KI per kilogram of diet. The iodine content of the basal diet determined by the ElmslieCaldwell method (A.O.A.C, 1960) was no greater than 0.3 p.p.m. iodide. The 0.3 p.p.m. level has been shown to be the minimum iodine requirement for normal thyroid function in Bobwhites (Kirk-
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Effects of Dietary Iodine on the Thyroidal Uptake and Elimination of 131-Iodine in Coturnix and Bobwhite Quail1
and treated. Holding birds for three days significantly reduced nine-day growth to a b o u t 9 0 % of t h a t of other holding treatments. This difference is probably due to the total number of days on feed, which indicates t h a t the earlier feeding h a d been most beneficial by adding extra days on feed. Mortality was slightly b u t not consistently highest in the control group.
157
tribute to early poult mortality in control, untreated groups. ACKNOWLEDGMENT T h e supply of water-soluble vitamins and neomycin was supplied through the courtesy of Nopco Chemical Company. Appreciation is also extended to Hoffmann-La Roche for their generous supply of vitamins for the basal diet.
SUMMARY REFERENCES Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Harper, J. A., and W. E. Babcock, 1953. The effect of penicillin on early mortality and growth in poults. Poultry Sci. 32: 179-180. Jensen, L. S., J. Allred, R. Fry and J. McGinnis, 1956. Effect of vitamin injections on survival of chicks with a high mortality syndrome. Fed. Proc. 15: 558. National Research Council, 1966. Nutrient Requirements of Domestic Animals. I. Nutrient Requirements of Poultry. National Academy of Science, Washington, D. C. Nestor, K. E., and K. I . Brown, 1966. Holding poults after hatching reduces early growth rate. Ohio Agri. Expt. Report 51: 36. Snedecor, G. W., and W. G. Cochran, 1967. Statistical Methods. Sixth edition. Iowa State University Press, Ames, Iowa. 593 pp.
Mycoplasma synoviae and Mycoplasma gallisepticum Infections: Differentiation by the HemagglutinationInhibition Test T. H. VARDAMAN 1 AND HARRY W. YODER, JR.2 United States Department of Agriculture, Animal Disease and Parasite Research Division (Received for publication August 18, 1969)
O
N E agent responsible for the clinical syndrome in chickens known as infectious synovitis, cultivated b y Chalquest and Fabricant (1960), was designated 1 South Central Poultry Research Laboratory, State College, Mississippi 39762. 2 Southeast Poultry Research Laboratory, Athens, Georgia 30601.
Mycoplasma synoviae by Olson et al. (1964). An important criterion for determining whether M. synoviae infection is present in a flock is the presence or absence of specific antibody as demonstrated by serologic examination. A M. synoviae serum plate agglutination test antigen has been produced (Olson et al.,
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 1, 2015
Turkey poults were pre-fed, when hatched or u p to three days after hatching, utilizing a technique of oral-forced pre-feeding of solutions as follows: plain tap water, vitamin, or vitamin plus neomycin. Then poults were given conventional diets and management, and monitored for 7 to 11 days. Significant increases in body weight usually resulted from pre-feeding the vitamin mixture or the vitamin plus neomycin solution as compared with no pre-feeding. A significant reduction in mortality during this period was recorded; mortality was usually lowest for those groups receiving the vitamin plus neomycin oral feeding. Dehydration appeared to substantially con-
158
T. H. VARDAMAN AND H. W. YODER, JR.
This study was done to determine if the M. synoviae and M. gallisepticum HI tests could be used to differentiate between M. synoviae and M. gallisepticum infections. MATERIALS AND METHODS
The chicken serums were obtained from birds infected by: 1) injecting isolate WVU 1853 or ISU isolate 1331 M. synoviae egg-yolk material into the hock joint or foot pad; 2) contact with birds infected with M. synoviae; 3) injecting M. gallisepticum egg-yolk into the airsacs; 4) contact with birds infected with M. gallisepticum; and 5) natural infection. The artificially infected chickens were grown in Horsfall and Bauer (1940) type isolation cabinets as modified by Drury et al. (1969). Serum samples from groups 1, 2, and 3 were from chickens inoculated at 3 weeks of age via foot pad or hock joint with 0.2 ml. of a 1:5, 1:25, or 1:100 dilution of M. synoviae egg-yolk. Serum
samples were obtained 5, 10, and 15 weeks postinoculation. Groups 4, 5, and 6 were serums from chickens that had been in contact with M. synoviae-mocvla.X.eA birds for 5, 10, and 15 weeks, respectively. Serum samples from group 9 were from turkeys that were inoculated at 5 weeks of age via air sac and sinus with 0.1 ml. of a 24-hour broth culture of M. synoviae strain 6130. Serum samples were obtained 5 weeks postinoculation. Serum samples from groups 11, 12, and 13 were from chickens inoculated at 3 weeks of age via air sacs with 0.2 ml. of M. gallisepticum undiluted egg-yolk. Serum samples were obtained 5, 8, and 11 weeks postinoculation. The methods of producing the M. synoviae hemagglutinating antigen and the procedure for the hemagglutinationinhibition test have been described (Vardaman and Yoder, 1969). The serums were tested by the HI test using M. synoviae and M. gallisepticum HA antigens, and the rapid serum plate test using M. synoviae serum plate test antigen and M. gallisepticum serum plate test antigen. The results of the HI and rapid serum plate agglutination tests were compared. RESULTS AND DISCUSSION
The serologic results from M. synoviaeand M. gallisepticum-miected birds are shown (Table 1). There were two different birds of the M. synoviae artificiallyinoculated group that had M. gallisepticum HI titers of 1:40 only. Bird 2012, from group 1, had an M. gallisepticum HI titer of 1:40 and a negative reaction to the M. gallisepticum serum plate test. It had an M. synoviae HI titer of 1:80 and a 3 + reaction to M. synoviae serum plate test. Bird 2077, from group 2, had an M. gallisepticum HI titer of 1:40 and a negative reaction to M. gallisepticum serum
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1963; and Roberts and Olesiuk, 1967). Olsen et al. (1964), and Roberts and Olesiuk (1967) reported that M. synoviaepositive serums would sometimes cause a serologic reaction with M. gallisepticum serum plate antigen. M. synoviae serum plate antigen is the only antigen commercially available for serologic examination for M. synoviae antibodies. Vardaman and Yoder (1969) reported the preparation of a M. .rywcwae-hemagglutinatmg antigen that showed good specificity when used in the hemagglutination-inhibition (HI) test. The hemagglutinationinhibition test is an official test, and is the most accurate serologically, for detecting antibodies against M. gallisepticum. It would seem desirable to have an HI test available for M. synoviae so that diagnostic laboratories could differentiate between M. synoviae and M. gallisepticum infections.
MYCOPLASMA
SYNOVIAE
159
AND GALLISEPTICUM
TABLE 1.—Results of survey of serologic differentiation between M. synoviae and M. gallisepticum
Group
li
chicken chicken chicken chicken chicken chicken chicken chicken turkey turkey chicken chicken chicken chicken turkey
Type infec.
Ms. Ms. Ms. Ms. Ms. Ms. Ms. Ms.+Mg. Ms. Ms. Mg. Mg. Mg. Mg. Mg.
Meth. infec.
Art. Art. Art. Cont. Cont. Cont. Field Field Art. Field Art. Art. Art. Cont. Cont.
Total No. birds 111 99 33 19 14 20 30 24 16 19 46 42 24 70 3
H I titer of 1 :40 & above M. synoviae %
Mean
82.9 94.9 93.9 21.1 78.6 90.0 83.3 91.7 68.8 78.9 0.0 0.0 0.0 0.0 0.0
238.3 291.9 238.7 110.0 138.2 248.8 134.4 225.5 225.5 304.0 0.0 0.0 0.0 0.0 0.0
M. gallisepticum % 0.9 1.01 0.0 0.0 0.0 5.0 0.0 87.5 0.0 0.0 100.0 100.0 100.0 100.0 100.0
Serum plate agglutination test—positive M. synoviae
Mean
%
40.0 40.0 0.0 00.0 00.0 40.0 00.0 81.9 00.0 00.0 219.1 310.5 246.7 421.7 266.7
98.20 98.90 100.00 42.11 92.86 85.00 86.66 100.00 25.00 100.00 58.70 16.67 4.17 0.00 66.67
No. 109 98 33 8 13 17 26 24 4 13/135 27 7 1 0 2
M. gallisepticum
% 0.00 0.00 12.10 0.00 0.00 0.00 83.33 95.83 0.00 69.20 95.65 100.00 100.00 94.30 100.00
No. 0 0 4 0 0 0 25 23 0 9/135 44 42 24 66 3
i2 Groups 1, 2, and 3 were the same birds, with serum samples obtained 5, 10, and 15 weeks postinoculation with M. synoviae. Groups 4, 5, and 6 were the same birds, with serum samples obtained 5,10, and 15 weeks post contact with M. synoviae-inoculated birds. 3 Group 9 were turkeys inoculated via sinus and trachea at 5 weeks, with serum samples obtained 5 weeks postinoculation. 4 Groups 11,12, and 13 were the same birds, with serum samples obtained 5,8, and 11 weeks postinoculation with M. gallisepticum. 5 Number positive over the number tested.
plate test, but it had an M. synoviae HI titer of 1:640 and a 3 + reaction to M. synoviae serum plate test. Group 2 of the M. synoviae artificially-inoculated birds, from which samples were obtained at 10 weeks postinoculation, had a higher percentage of M. synoviae H I titers of 1:40 and a higher mean than groups 1 and 3 from which serum samples were obtained at 5 and 15 weeks postinoculation, respectively. The mean M. synoviae H I titers were 238.3,291.9, and 238.7 from groups 1, 2, and 3, respectively. The lower percentage of M. synoviae H I positive results and the mean M. synoviae HI titers of group 3, when compared to group 2, are due to the fact that most of the obviously sick birds either died or were killed before 15 weeks. Typical reactions of M. synoviae artificially-inoculated birds were two which had M. synoviae HI titers of 1:40, 1:80, and 1:320 at 5, 10, and 15 weeks postinoculation, respectively. Bird 1608, from group 6, had an M. gallisepticum HI titer of 1:40 and a negative reaction to the M. gallisepticum serum plate test, but it had an M. synoviae HI titer of 1:640 and a 4 + reaction to M.
synoviae serum plate test. The percentage of M. synoviae HI titers of 1:40 and above, and the mean HI titers of the M. synoviae contact birds increased with time as shown in Table 1, by comparing groups 4, 5, and 6. Group 7, field samples of M. synoviae suspects, did not show any reactions to M. gallisepticum HI test and had a mean M. synoviae H I titer of 134.4. There were many M. gallisepticum serum plate crossreactions since these samples were selected to confirm M. synoviae infection by using the HI test. Group 8 were field samples that had positive reactions to both the M. synoviae and M. gallisepticum HI test and serum plate tests. This flock had been diagnosed as having M. synoviae plate- and Hl-positive reactions and M. gallisepticum plateand Hi-negative reactions previously. Three months later, when the 24 serum samples were taken, this flock began to show respiratory signs. Serologic results at this time showed that the flock was infected with M. gallisepticum and M. synoviae. Group 9 were serum samples from tur-
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21 31 42 52 62 7 8 93 10 11« 12* 13« 14 15
Species
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T . H . VARDAMAN AND H . W. Y O D E R , J R .
keys t h a t had been inoculated with a broth culture of M. synoviae via sinus and trachea 5 weeks previously. Approximately 6 9 % of the birds had a M. synoviae H I titer of 1:40 and above with a mean titer of 225.5. Only 25.0% h a d a reaction to the M. synoviae serum plate test and they had 1 + reactions or very slight reactions. No turkey serums reacted to the M. gallisepticum H I test or serum plate test.
T h e d a t a show considerable cross reactions to M. synoviae and M. gallisepticum serum plate tests in field samples from chickens and turkeys. T h e y show a need for the M. synoviae H I test which, used in conjunction with the M. gallisepticum H I test, would aid the diagnostician in differentiating between M. synoviae and M. gallisepticum infections.
Group 10 were selected turkey serums from the field flocks sent to the laboratory for confirmation of results. They had strong M. synoviae serum plate reactions and h a d a mean M. synoviae H I titer of 304.0. The M. gallisepticum serum plate reactions were weak, a 1 + or 2 + , and all were negative to M. gallisepticum HI test. Groups 11, 12, and 13 were artificially infected with undiluted M. gallisepticum egg-yolk material b y injecting the material into the air sacs. T h e y were all positive to the M. gallisepticum H I test with a mean H I titer of 219.1, 310.5, and 246.7 at 5, 8, and 11 weeks postinoculation. There were no cross reactions to M. synoviae H I test. There were 58.7, 16.7, and 4 . 2 % reactors to the M, synoviae serum plate test compared to 95.6, 100.0, and 100.0% reactors to M. gallisepticum serum plate test, a t 5, 8, and 11 weeks postinoculation. The initial cross reactions to the M. synoviae serum plate test m a y be caused b y heavy yolk inoculation to infect the birds.
T h e hemagglutination-inhibition tests for Mycoplasma synoviae and M. gallisepticum were used on serum samples from artificially-infected, contact-exposed and normal chickens raised in isolation cabinets. Other samples from field flocks of chickens and turkeys were also tested. Antigens prepared for both M. synoviae a n d M. gallisepticum had good specificity in differentiating between these infections by the hemagglutination-inhibition procedure. Cross reactions detected by the rapid serum plate test procedure were not a problem in the H I test. T h e tests worked equally well with both chicken and turkey serums, and should be valuable for diagnostic purposes.
Group 14, M. gallisepticum contact chickens, reacted to M. gallisepticum H I and serum plate test only. Group 15, M. gallisepticum contact turkeys, show M. gallisepticum H I and serum plate positive reactions. They had negative results to M. synoviae H I test but some reacted to M. synoviae serum plate test.
Chalquest, R. R., and J. Fabricant, 1960. Pleuropneumonia-like organisms associated with synovitis in fowls. Avian Diseases, 4: 515-539. Drury, L. N., C. W. Beard and S. R. Hopkins, 1969. An isolation cabinet system for avian disease research. Avian Diseases, 13: 400-412. Horsfall, F. L., Jr., and J. H. Bauer, 1940. Individual isolation of infected animals in a single room. J. Bact. 40: 569-580. Olson, N. O., K. M. Kerr and A. Campbell, 1963. Control of infectious synovitis. 12. Preparation
SUMMARY
T h e authors express their appreciation to Mrs. Charmian Lee Gatlin a n d Mr. Boyd Turner, medical technologists, for their assistance. REFERENCES
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ACKNOWLEDGEMENTS
MYCOPLASMA SYNOVIAE AND GALLISEPTICUM of an agglutination test antigen. Avian Diseases, 7: 310-317. Olson, N. O., K. M. Kerr and A. Campbell, 1964. Control of infectious synovitis. 13. The antigen study of three strains. Avian Diseases, 8: 209214. Roberts, D. H., and 0. M. Olesiuk, 1967. Serological
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studies with Mycoplasma synoviae. Avian Diseases, 11: 104-119. Vardaman, T. H., and H. W. Yoder, Jr., 1969. Preparation of Mycoplasma synoviae hemagglutinating antigen and its use in the hemagglutinationinhibition test. Avian Diseases, 13: 654-661.
ERIC L. MORGAN2 AND FRANK R. MRAZ Agricultural Research Laboratory of the University of Tennessee, Oak Ridge, Tennessee 37830 (Received for publication August 18, 1969)
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ARIATIONS in time and level of maximum thyroidal uptake or biological half-life of 181I following a single dose have been reported in several species of birds (McFarland et al., 1966; Rosenberg et al., 1963a; Premachandra et al., 1958; Wahlberg, 1955). Featherston et al. (1966) found that chicks fed an iodinedeficient diet retained more 131I originally than did control chicks. Thyroids of iodine-deficient chicks trapped a maximum of 4 1 % of the original dose at 2 hr. with a biological half-life of 15 hr. Control chicks, however, trapped a maximum of only 4.5% at 8 hr with a biological half-life of more than 96 hr. The primary objectives of this study were to determine the effects of dietary iodine on the maximum uptake and biological half-life of 131I in the thyroid glands of Coturnix and Bobwhite quail and to 1 This manuscript is published with the permission of the Director of The University of Tennessee Agricultural Experiment Station, Knoxville, Tennessee. 2 Present Address: Department of Biology, The University of Tennessee, Chattanooga, Tennessee. 3 Operated by the Tennessee Agricultural Experiment Station for the U. S. Atomic Energy Commission under Contract No. AT-40-1-GEN-242.
compare the response of these two species with respect to thyroidal 13lI metabolism. EXPERIMENTAL All quail used in these studies were hatched from breeding stocks at the UTAEC Agricultural Research Laboratory. The Coturnix were originally acquired from Auburn University and the Bobwhites from the Tennessee Wildlife Commission's Buffalo Springs Game Bird Farm. Day-old Coturnix and Bobwhite chicks were placed in specially constructed 1/4-inch wire mesh screen cages adapted to jfelectrically-heated commercial chick brooders. They were immediately fed the experimental diets and at about 2 weeks of age moved to standard chick brooders. Half the birds in each experiment were fed the pheasant and quail starter diet of Scott et al. (1960) minus the Ca(IOs)2. The remaining half were fed this diet supplemented with 0.1 mg. iodide from KI per kilogram of diet. The iodine content of the basal diet determined by the ElmslieCaldwell method (A.O.A.C, 1960) was no greater than 0.3 p.p.m. iodide. The 0.3 p.p.m. level has been shown to be the minimum iodine requirement for normal thyroid function in Bobwhites (Kirk-
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Effects of Dietary Iodine on the Thyroidal Uptake and Elimination of 131-Iodine in Coturnix and Bobwhite Quail1