The Ability of Blood Plasma from Drinking Water Vaccinated Turkeys to Protect Against a Lethal Challenge of Pasteurella multocida B. W. BlERER AND W. T. DERIEUX South Carolina Agricultural Experiment Station, Clemson University, Clemson, South Carolina 29631* (Received for publication March 31, 1975)
POULTRY SCIENCE 54: 2091-2093, 1975
encourage prompt consumption of the inoculated water, the drinking water was withheld AHESWARAN et al. (1973) have 2 hours prior to the exposure to the vaccine. reported that their Pasteurella multociA separate flask of 24-hours old broth culture da mutant (M-2283) strain, used as a drinking was used on each of the second, third and water vaccine, resulted in a localized protecfourth days. tion but did not provide systemic protection. Three weeks after vaccination, 10 ml. of It was the purpose of the work reported to heparinized blood was obtained from each determine whether or not the Clemson Uniturkey and identified as to source. Samples versity (C.U.) strain (Bierer and Derieux, were also obtained from 4 nonvaccinated 1972) was capable of providing systemic proturkeys in a separate chamber. Plasma was tection when used as a drinking water vacharvested from each sample separately and cine. frozen. These turkeys were then exposed to a virulent P-1059 strain of P. multocida, using MATERIALS AND METHODS the drinking water route and using the same Sixteen 15-weeks-old turkeys were vacci- procedure described in the foregoing for nated against fowl cholera disease with the vaccination. C.U. vaccine strain, using the water route. Eight of the 16frozen plasma samples, from The vaccination procedure has been de- the turkeys that had been vaccinated, were scribed (Bierer and Derieux, 1972) and in- thawed and 0.3 ml. of plasma from each cludes the addition of 30 ml. of a 24-hours sample was injected into each of 10 6-day old brain-heart-infusion broth culture (incu- old baby chicks. The plasma was injected bated at 37 degrees C.) to each 4,000 ml. into the subcutaneous tissue between the of drinking water for 4 consecutive days. To thigh and abdominal cavity. At the same time each of these chicks was also injected into the breast muscle with 0.1 ml. of a 10~3 *Published with approval of the Director as Technidilution of a 24-hours old brain-heart-infusion cal Contribution No. 1249. INTRODUCTION
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ABSTRACT Turkeys were vaccinated against fowl cholera with the Clemson University avirulent Pasteurella multocida vaccine strain (C.U. strain), using the drinking water route. Plasma was harvested from these turkeys, which later withstood challenge with a highly virulent P-1059 strain of P. multocida. Baby chicks were injected between the thigh and abdominal cavity with the immune plasma and simultaneously, into the breast muscle, with the same highly virulent strain referred to in the foregoing. The results demonstrated that plasma from turkeys vaccinated by the drinking water route protected baby chicks against intramuscular challenge with a virulent strain of P. multocida and suggest that a favorable systemic immunologic response did occur. This is in contrast to the Minnesota M-2283 strain which, reportedly, produces a localized protection only, when used as a drinking water vaccine.
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ples from nonvaccinated and nonimmunized turkeys. The daily mortality record of the chicks that were injected with plasma and then simultaneously injected with a virulent strain of Pasteurella is given in Table 1. The average number of deaths per group in each brooder was less in those groups that had received the plasma from the vaccinated turkeys when compared to the deaths that occurred in the groups that had received normal plasma from nonvaccinated turkeys or in the groups receiving no plasma at all. The results, submitted to a statistician (Birdsall, 1973) were evaluated, as follows: "There is no good way to analyze this data statistically but there is no doubt that (a) there is no significant difference between normal and no serum relative to 7 day chick mortality and (b) the immune serum has a significant effect in reducing 7 day chick mortality. This treatment is significantly better than either no serum or normal serum."
RESULTS AND DISCUSSION The 16 turkeys vaccinated with the C.U. strain all survived challenge when exposed to the virulent P-1059 P. multocida strain and all 4 of the nonvaccinated turkeys succumbed to the challenge. These results suggest that the plasma samples obtained from the vaccinates were representative samples from immunized turkeys and that the samples from the nonvaccinates were representative sam-
Maheswaran et al. (1973) have reported that a live P. multocida mutant used as a live drinking water vaccine in turkeys produced a localized protection in the respiratory sys-
TABLE 1.—The ability of blood plasma from turkeys that had received C.U. strain Pasteurella multocida drinking water vaccine to protect baby chicks against virulent P-1059 strain challenge
1 2 3 4 5 6 7 8 9 10 11 12
Treatment
Number of chicks
1
Deaths—days post challenge 2 3 4 5 6
Immune plasma Immune plasma Immune plasma Immune plasma Normal plasma No plasma Immune plasma Immune plasma Immune plasma Immune plasma Normal plasma No plasma
10 10 10 10 10 10 10 10 10 10 8 10
1 1 2 0 4 6 0 1 1 0 4 8
1 0 4 0 3 2 3 2 0 5 2 1
*Six days old White Leghorn Cockerels. **The number that died over the number of chicks.
0 2 0 0 1 1 0 0 1 0 1 0
0 1 0 1 1 0 1 1 0 0 0 0
0 0 1 1 0 0 0 0 1 0 0 0
0 0 0 0 0 0 0 0 0 0 0 0
Total died** 0 0 0 0 0 0 0 0 0 0 0 0
2/10 4/10 7/10 2/10 9/10 9/10 4/10 4/10 3/10 5/10 7/8 9/10
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broth culture of a virulent P-1059 P. multocida strain. The results of a plate count technique (Heddleston, 1972) indicated that approximately 119,000 viable bacilli were injected into each chick. Each group of 10 chicks was placed into separate chick brooders in the experimental design. Mortality data was recorded daily. Two additional groups of 10 chicks were injected with 0.3 ml. of plasma from each of two plasma samples from turkeys that had not been vaccinated. Two additional groups of 10 were injected with 0.3 ml. of sterile saline. The chicks in each of these 4 additional groups were also injected into the breast muscle with 0.1 ml. of the P-1059 virulent culture referred to in the previous paragraph.
PASTEURELLA
While the results of the work reported in this present paper suggest that a systemic protection occurred after the use of the C.U. strain as a drinking water vaccine in turkeys, these results certainly do not refute the possibility or even the probability that a localized protection in the respiratory system also did occur, as well. Gomez and Raggi (1974) observed a local resistance to the infectious bronchitis virus in trachael organ culture from chickens previously immunized against the infectious bronchitis virus. It appears (Baron, 1963) that in virus infections this local protective mechanism is capable of working either alone or in cooperation with the general systemic immune mechanism. Maheswaran et al. (1973), working with their P. multocida
M-2283 strain, noted that the mechanism of the local immunity observed was unknown and suggested that the local immunity could be the result of either humoral antibody, or of cell mediated immunity or both. ACKNOWLEDGEMENTS Technical details relative to the evaluation of the plasma samples referred to in this paper were kindly provided by Kenneth L. Heddleston (Senior Microbiologist, National Animal Disease Laboratory, Ames, Iowa 50010), who also provided the pathogenic P-1059 strain of P. multocida. REFERENCES Baron, S., 1963. Mechanisms of recovery from viral infection. Adv. Virus Res. 10: 39-64. Bierer, B. W., and W. T. Derieux, 1972. Immunologic response of turkeys to an avirulent Pasteurella multocida vaccine in the drinking water. Poultry Sci. 51: 408-416. Birdsall, J. J., 1973. Official communication, May 21, 1973, WARF Institute, Inc., P.O. Box 3599, Madison, Wisconsin 53701. Gomez, L., and L. G. Raggi, 1974. Local immunity to avian infectious bronchitis in tracheal organ culture. Avian Dis. 8: 346-368. Heddleston, K. L., 1972. Personal communication, January 24. Maheswaran, S. K., J. R. McDowell and B. S. Pomeroy, 1973. Studies on Pasteurella multocida. I. Efficacy of an avirulent mutant as a live vaccine in turkeys. Avian Dis. 17: 396-405.
NEWS AND NOTES (Continued from page 2090) in the three official languages. 10. The complete text of reviews and papers will be printed in one of the official languages and published with the abstracts in the Proceedings of the Conference, shortly after Conference. 11. Any member who proposes to submit a paper must apply for the official forms and typing
instructions to the Conference Secretariat: Mr. J. R. Debattista, Director of Conference, 1, Simon Flats, Dr. Zammit Street, Balzan, Malta. 12. During four afternoons, six symposia will be held on special topics without simultaneous translation. These topics are: (a) geese and ducks, (b) turkeys, (c) the effect of energy costs on the reuse or
(Continued on page 2114)
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tem, but did not provide a systemic protection. The belief of these workers was based on their observation that turkeys vaccinated by the oral or by the endotracheal routes succumbed when challenged by an intramuscular injection of virulent P. multocida bacilli. Unfortunately, the isolate used by Maheswaran et al. (1973) (designated as M2283) is not available for evaluation by workers outside of the State of Minnesota. Perhaps, in the hands of other workers and the use of varied techniques of evaluation, certain workers would produce evidence that the M-2283 strain used by Maheswaran et al. (1973), when used orally, does elicit a degree of systemic protection.
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MULTOCIDA
POULTRY SCIENCE 54: 2091-2093, 1975
encourage prompt consumption of the inoculated water, the drinking water was withheld AHESWARAN et al. (1973) have 2 hours prior to the exposure to the vaccine. reported that their Pasteurella multociA separate flask of 24-hours old broth culture da mutant (M-2283) strain, used as a drinking was used on each of the second, third and water vaccine, resulted in a localized protecfourth days. tion but did not provide systemic protection. Three weeks after vaccination, 10 ml. of It was the purpose of the work reported to heparinized blood was obtained from each determine whether or not the Clemson Uniturkey and identified as to source. Samples versity (C.U.) strain (Bierer and Derieux, were also obtained from 4 nonvaccinated 1972) was capable of providing systemic proturkeys in a separate chamber. Plasma was tection when used as a drinking water vacharvested from each sample separately and cine. frozen. These turkeys were then exposed to a virulent P-1059 strain of P. multocida, using MATERIALS AND METHODS the drinking water route and using the same Sixteen 15-weeks-old turkeys were vacci- procedure described in the foregoing for nated against fowl cholera disease with the vaccination. C.U. vaccine strain, using the water route. Eight of the 16frozen plasma samples, from The vaccination procedure has been de- the turkeys that had been vaccinated, were scribed (Bierer and Derieux, 1972) and in- thawed and 0.3 ml. of plasma from each cludes the addition of 30 ml. of a 24-hours sample was injected into each of 10 6-day old brain-heart-infusion broth culture (incu- old baby chicks. The plasma was injected bated at 37 degrees C.) to each 4,000 ml. into the subcutaneous tissue between the of drinking water for 4 consecutive days. To thigh and abdominal cavity. At the same time each of these chicks was also injected into the breast muscle with 0.1 ml. of a 10~3 *Published with approval of the Director as Technidilution of a 24-hours old brain-heart-infusion cal Contribution No. 1249. INTRODUCTION
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ABSTRACT Turkeys were vaccinated against fowl cholera with the Clemson University avirulent Pasteurella multocida vaccine strain (C.U. strain), using the drinking water route. Plasma was harvested from these turkeys, which later withstood challenge with a highly virulent P-1059 strain of P. multocida. Baby chicks were injected between the thigh and abdominal cavity with the immune plasma and simultaneously, into the breast muscle, with the same highly virulent strain referred to in the foregoing. The results demonstrated that plasma from turkeys vaccinated by the drinking water route protected baby chicks against intramuscular challenge with a virulent strain of P. multocida and suggest that a favorable systemic immunologic response did occur. This is in contrast to the Minnesota M-2283 strain which, reportedly, produces a localized protection only, when used as a drinking water vaccine.
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B . W . BlERER AND W . T . DERIEUX
ples from nonvaccinated and nonimmunized turkeys. The daily mortality record of the chicks that were injected with plasma and then simultaneously injected with a virulent strain of Pasteurella is given in Table 1. The average number of deaths per group in each brooder was less in those groups that had received the plasma from the vaccinated turkeys when compared to the deaths that occurred in the groups that had received normal plasma from nonvaccinated turkeys or in the groups receiving no plasma at all. The results, submitted to a statistician (Birdsall, 1973) were evaluated, as follows: "There is no good way to analyze this data statistically but there is no doubt that (a) there is no significant difference between normal and no serum relative to 7 day chick mortality and (b) the immune serum has a significant effect in reducing 7 day chick mortality. This treatment is significantly better than either no serum or normal serum."
RESULTS AND DISCUSSION The 16 turkeys vaccinated with the C.U. strain all survived challenge when exposed to the virulent P-1059 P. multocida strain and all 4 of the nonvaccinated turkeys succumbed to the challenge. These results suggest that the plasma samples obtained from the vaccinates were representative samples from immunized turkeys and that the samples from the nonvaccinates were representative sam-
Maheswaran et al. (1973) have reported that a live P. multocida mutant used as a live drinking water vaccine in turkeys produced a localized protection in the respiratory sys-
TABLE 1.—The ability of blood plasma from turkeys that had received C.U. strain Pasteurella multocida drinking water vaccine to protect baby chicks against virulent P-1059 strain challenge
1 2 3 4 5 6 7 8 9 10 11 12
Treatment
Number of chicks
1
Deaths—days post challenge 2 3 4 5 6
Immune plasma Immune plasma Immune plasma Immune plasma Normal plasma No plasma Immune plasma Immune plasma Immune plasma Immune plasma Normal plasma No plasma
10 10 10 10 10 10 10 10 10 10 8 10
1 1 2 0 4 6 0 1 1 0 4 8
1 0 4 0 3 2 3 2 0 5 2 1
*Six days old White Leghorn Cockerels. **The number that died over the number of chicks.
0 2 0 0 1 1 0 0 1 0 1 0
0 1 0 1 1 0 1 1 0 0 0 0
0 0 1 1 0 0 0 0 1 0 0 0
0 0 0 0 0 0 0 0 0 0 0 0
Total died** 0 0 0 0 0 0 0 0 0 0 0 0
2/10 4/10 7/10 2/10 9/10 9/10 4/10 4/10 3/10 5/10 7/8 9/10
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on May 24, 2015
broth culture of a virulent P-1059 P. multocida strain. The results of a plate count technique (Heddleston, 1972) indicated that approximately 119,000 viable bacilli were injected into each chick. Each group of 10 chicks was placed into separate chick brooders in the experimental design. Mortality data was recorded daily. Two additional groups of 10 chicks were injected with 0.3 ml. of plasma from each of two plasma samples from turkeys that had not been vaccinated. Two additional groups of 10 were injected with 0.3 ml. of sterile saline. The chicks in each of these 4 additional groups were also injected into the breast muscle with 0.1 ml. of the P-1059 virulent culture referred to in the previous paragraph.
PASTEURELLA
While the results of the work reported in this present paper suggest that a systemic protection occurred after the use of the C.U. strain as a drinking water vaccine in turkeys, these results certainly do not refute the possibility or even the probability that a localized protection in the respiratory system also did occur, as well. Gomez and Raggi (1974) observed a local resistance to the infectious bronchitis virus in trachael organ culture from chickens previously immunized against the infectious bronchitis virus. It appears (Baron, 1963) that in virus infections this local protective mechanism is capable of working either alone or in cooperation with the general systemic immune mechanism. Maheswaran et al. (1973), working with their P. multocida
M-2283 strain, noted that the mechanism of the local immunity observed was unknown and suggested that the local immunity could be the result of either humoral antibody, or of cell mediated immunity or both. ACKNOWLEDGEMENTS Technical details relative to the evaluation of the plasma samples referred to in this paper were kindly provided by Kenneth L. Heddleston (Senior Microbiologist, National Animal Disease Laboratory, Ames, Iowa 50010), who also provided the pathogenic P-1059 strain of P. multocida. REFERENCES Baron, S., 1963. Mechanisms of recovery from viral infection. Adv. Virus Res. 10: 39-64. Bierer, B. W., and W. T. Derieux, 1972. Immunologic response of turkeys to an avirulent Pasteurella multocida vaccine in the drinking water. Poultry Sci. 51: 408-416. Birdsall, J. J., 1973. Official communication, May 21, 1973, WARF Institute, Inc., P.O. Box 3599, Madison, Wisconsin 53701. Gomez, L., and L. G. Raggi, 1974. Local immunity to avian infectious bronchitis in tracheal organ culture. Avian Dis. 8: 346-368. Heddleston, K. L., 1972. Personal communication, January 24. Maheswaran, S. K., J. R. McDowell and B. S. Pomeroy, 1973. Studies on Pasteurella multocida. I. Efficacy of an avirulent mutant as a live vaccine in turkeys. Avian Dis. 17: 396-405.
NEWS AND NOTES (Continued from page 2090) in the three official languages. 10. The complete text of reviews and papers will be printed in one of the official languages and published with the abstracts in the Proceedings of the Conference, shortly after Conference. 11. Any member who proposes to submit a paper must apply for the official forms and typing
instructions to the Conference Secretariat: Mr. J. R. Debattista, Director of Conference, 1, Simon Flats, Dr. Zammit Street, Balzan, Malta. 12. During four afternoons, six symposia will be held on special topics without simultaneous translation. These topics are: (a) geese and ducks, (b) turkeys, (c) the effect of energy costs on the reuse or
(Continued on page 2114)
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on May 24, 2015
tem, but did not provide a systemic protection. The belief of these workers was based on their observation that turkeys vaccinated by the oral or by the endotracheal routes succumbed when challenged by an intramuscular injection of virulent P. multocida bacilli. Unfortunately, the isolate used by Maheswaran et al. (1973) (designated as M2283) is not available for evaluation by workers outside of the State of Minnesota. Perhaps, in the hands of other workers and the use of varied techniques of evaluation, certain workers would produce evidence that the M-2283 strain used by Maheswaran et al. (1973), when used orally, does elicit a degree of systemic protection.
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MULTOCIDA
