Immunologic Response of Turkey Poults of Various Ages to an Avirulent Pasteurella multocida Vaccine in the Drinking Water* B . W. BlERER AND W. T. DERIEUX
South Carolina Agricultural Experiment Station, Clemson University, Clemson, South Carolina 29631 (Received for publication August 23, 1974)
POULTRY SCIENCE 54: 784-787, 1975
INTRODUCTION INCE the published evaluation of an avirulent Pasteurella multocida drinking water vaccine by Bierer and Derieux (1972), this type vaccine has been used rather extensively by commercial turkey producers in the Carolinas (Derieux, 1974) and is referred to as the C.U. (Clemson University) strain drinking water vaccine. The usual procedure in the field has been to administer an injectable oil base P. multocida bacterin to each turkey at 8 weeks of age and then to give the drinking water vaccine three to four weeks later. The Clemson Livestock Laboratory at Columbia, South Carolina reported 5 outbreaks of fowl cholera in turkeys during 1973 alone that occurred around 5 to 6 weeks of age
and prior to the initiation of a fowl cholera control program at 8 weeks of age. In view of this, an experiment was conducted to obtain information relative to the immunologic response of small turkey poults to an avirulent P. multocida vaccine administered in the drinking water.
S
*Published with approval of the Director as Technical Contribution No. 1204.
MATERIALS AND METHODS One hundred and sixty baby turkey poults were obtained from a commercial turkey hatchery. These poults had not been injected with any type of medication or vaccine and had not been sexed. According to hatchery records the poults were the progeny of a breeder flock that had been on a fowl cholera control program that included the use of the C.U. strain drinking water vaccine prior to the time the breeders came into production. The vaccine strain has been described in a previous paper (Bierer and Derieux, 1972). Twenty of the day old poults were placed
784
Downloaded from http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
ABSTRACT Fowl cholera has been reported in turkey poults in South Carolina between 5 and 6 weeks of age. An experiment was conducted to determine the earliest age that poults could be vaccinated with the Clemson University Pasteurella multocida strain (using the drinking water route) that would result in an effective immunity at 5 to 6 weeks of age. In groups of vaccinates challenged with a virulent P. multocida (P-1059 strain) at 5 weeks of age, none were infected in groups vaccinated at either 20 days or at 25 days, 1 of 10 vaccinated at 15 days, 3 of 10 vaccinated at 10 days, 6 of 10 vaccinated at 5 days, and 8 of 10 became infected of those vaccinated at 1 day of age. Eight of 10 of each of 2 groups of nonvaccinates developed evidence of infection. In groups of vaccinated challenged at 12 weeks of age, 2 of 10 turkeys became infected that had been vaccinated at 25 days of age, 4 of 10 that had been vaccinated at 20 days, 8 of 10 vaccinated at 15 days, 8 of 10 vaccinated at 10 days, 8 of 9 vaccinated at 5 days, and 8 of 9 became infected of those vaccinated at 1 day of age. All 10 of one group of nonvaccinates and all 9 of a second group developed evidence of infection. These results suggest, under the conditions of this experiment, that turkey poults vaccinated at 35 days of age and poults vaccinated at 20 days of age experienced an excellent immunological response by 5 weeks of age. This response was less evident but, for the most part, still present, at 12 weeks of age.
PASTEURELLA
MULTOCIDA VACCINE
At 3 weeks of age, each group of poults was moved to separate growing units. 2 At 5 weeks of age, half of each of the 8 groups were placed in each of 8 individual floor pen units. The following day the poults remaining in the growing units were exposed to a highly virulent P. multocida (P-1059 strain) strain obtained from Kenneth Heddleston, Principal Research Microbiologist at the National Animal Disease Laboratory at Ames, Iowa. All 8 of the groups in floor pens were exposed to the highly virulent P. multocida strain at 12 weeks of age. Exposure, in each instance, was accomplished by the drinking water route, using a 1:50 dilution of a 20
1. Petersime Brooder Units, Model 2SD modified to single deck units. Petersime Incubator Company, Gettysburg, Ohio 45328. 2. Petersime Finishing Unit No. 4 modified to single deck units. This is a unit designed for turkey research manufactured by the Petersime Incubator Company, Gettysburg, Ohio 45328.
hours old brain-heart infusion broth culture as the only source of drinking water daily, for 3 consecutive days. Drinking water was not withheld prior to the use of this culture. Observations on all groups exposed to the highly virulent strain were discontinued 2 weeks after such exposure. Mortality data was recorded daily and the number morbid at termination was recorded. Mortality, plus morbidity was then recorded as the total number infected. The experimental facilities were housed in a well constructed old dairy barn that had been converted into areas for poultry research. The 8 floor pen units, used to house the eight treatment groups after 5 weeks of age, were of concrete block construction with a separate entrance into an anteroom. The anteroom contained feed and water facilities for the adjacent chambers and each chamber was 2.7 x 3.4 meters. These units were routinely cleaned and disinfected and the chambers prepared with fresh, clean shavings litter prior to their use. There were no other disease experiments in progress on the immediate premises. All groups were maintained in a manner that was designed to prevent direct contact of the birds between the different treatment groups. RESULTS AND DISCUSSION The results of the challenge with the highly virulent P. multocida strain at 5 weeks of age are given in Table 1. All 20 of the turkeys vaccinated at 25 and at 20 days of age survived this challenge and remained clinically healthy during the 2 weeks post challenge period of observation. Evidence of infection appeared in groups vaccinated at 15 days, 10 days, 5 days and at 1 day. The infection rate, in these groups, increased from 1 turkey in the 10 vaccinated at 15 days of age to 8 of the 10 turkeys vaccinated at 1 day of age. The results of the challenge with the virulent P. multocida strain at 12 weeks of age are given in Table 2. Fowl cholera infection
Downloaded from http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
in each of 8 brooder units. 1 C.U. strain drinking water vaccine was administered in the drinking water to the poults in the first group during the first day of age and daily thereafter, for a total of 3 consecutive days, as the only source of drinking water. The vaccine mixture was a 1:50 dilution of a 20 hours old brainheart infusion broth culture of the C.U. strain. A tenfold dilution plate count technique (Heddleston, 1972) on dextrose starch agar established that each ml. of water vaccine contained approximately 20,000,000 viable bacilli when prepared. The second group of poults received the vaccine at 5 days of age, the third at 10 days, the fourth at 15 days, the fifth at 20 days, and the sixth group of poults at 25 days of age. In each instance, as with the first group, the vaccine was given as the only source of drinking water daily, for 3 consecutive days. Drinking water was not withheld prior to the use of the vaccine. The 20 poults in each of the seventh and eighth groups were not vaccinated.
785
— —
Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Nonvaccinated Nonvaccinated
Treatment status
— —
Age when vaccinated (days) 1 5 10 15 20 25
*Number affected over number of turkeys.
Units 1 2 3 4 5 6 7 8
2 2 1 0 0 0 0 2 3 3 1 1 1 0 0 0 1 1 4 0 2 0 0 0 0 1 0
Number died (days postchallenge) 5 6 7 8 1 1 1 0 1 1 0 0 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 3 1 0 0 9 0 0 0 0 0 0 1 0
10 0 0 0 0 0 0 1 0
2 4 5 4 2 0 0 8 9
3 3 1 3 2 0 0 0 0
Number died (days postchallenge) 5 6 4 0 0 1 0 2 0 1 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 1 0
10 0 0 0 1 0 0 0 0
TABLE 2.—Postvaccination results of challenge at twelve weeks of age with a virulent P-1059
*Number affected over number of turkeys.
Unit 1 2 3 4 5 6 7 8
Treatment status Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Nonvaccinated Nonvaccinated
Age when vaccinated (days) 1 5 10 15 20 25
TABLE 1.—Postvaccination results of challenge at five weeks of age with a virulent P-1059
om http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
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may make them poor subjects to vaccinate, with any type of vaccine, when a lasting immunological response is desired. The fact that the poults used in the trial described in this present paper could be, for the most part, successfully vaccinated at 25 and at 20 days of age and less successfully at 15, 10, and 5 days and at 1 day of age, suggests that some or all of the detrimental factors mentioned in this paragraph may have been at play. The reader must be reminded that it was not the objective of this experiment to determine the earliest age at which an effective immunological response would occur but, rather, the earliest age at which poults could be vaccinated that would result in an effective immunity at 5 or 6 weeks of age. The results do suggest, however, that if an immunological response did result from the day old or 5 days of age vaccinations, that there was little or no evidence of this response when challenged at 35 days (5 weeks) of age. REFERENCES Bierer, B. W.,and W. T. Derieux, 1972. Immunological response in turkeys to an avirulent Pasteurella multocida vaccine in the drinking water. Poultry Sci. 51:408-416. Bierer, B. W., T. H. Eleazer and D. E. Roebuck, 1964. Sedimentation rate, packed cell volume, buffy coat value, and rectal temperature of chickens and turkeys of various ages. J. Amer. Vet. Med. Ass. 144: 727-730. Derieux, W. T., 1974. Fowl cholera, is there hope for oral vaccine? Poultry Digest, March: 126-127. Heddleston, K. L., 1972. Personal communication, January 24, 1972. Jungherr, E., and N. Terrell, 1946. Observation of the spread of Newcastle disease. Proceedings of the 50th Annual Meeting of the U.S. Livestock Sanitary Ass., p. 170. Jungherr, E., and N. Terrell, 1948. Naturally acquired passive immunity to infectious bronchitis in chicks. Amer. J. Vet. Res. 9: 201. Shoffner, R. N., 1965. Heredity and the defective in poultry, age. In: Diseases of Poultry, Biester and Schwarte Ed., Iowa State University Press, Ames, Iowa 50010, 5th Ed., page 93.
Downloaded from http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
was observed in 2 of the 10 turkeys vaccinated at 25 days of age and in 4 of the 10 turkeys vaccinated at 20 days of age. Details relative to the challenge at 5 weeks of age and the challenge at 12 weeks of age are given in Table 1 and Table 2. During the first 5 days of age there was 1 death in the group vaccinated at 1 day and at 5 days, and 1 death in a nonvaccinated group. A bacteriological examination supplied evidence that these deaths were due to E. coli infection associated with oomphalitis. These were the only deaths during the course of this experiment other than those resulting from challenge with the virulent strain of P. multocida. The poults used in this trial were from parent stock that had been vaccinated at 27 weeks of age with the C.U. strain of Pasteurella multocida by the drinking water route. The transfer of passive immunity has been reported to occur from parent stock to offspring and to persist for as long as 4 weeks in Newcastle disease (Jungherr and Terrel, 1946) and with infectious bronchitis in chickens (Jungherr and Terrell, 1948), for example. Presumably, the presence of passive immunity to fowl cholera in baby poults could interfere with their successful vaccination by the drinking water route as described under materials and methods. Also, other factors may have contributed to the high infection rates resulting from the challenge applied to the poults vaccinated at 1 day and at 5 days of age (See Tables 1 and 2). Shoffner (1965) states that, "the embryonic chick and newly hatched chick are immunologically incompetent." Presumably, this same line of reasoning can be applied to the embryonic and newly hatched poult. Bierer et al. (1964) found that such values as sedimentation rate, erythrocyte volume, buffy coat and body temperatures were not stabilized in chicks and young turkeys until the tenth to fourteenth day of life. Thus, the physiological instability of baby chicks and baby poults
South Carolina Agricultural Experiment Station, Clemson University, Clemson, South Carolina 29631 (Received for publication August 23, 1974)
POULTRY SCIENCE 54: 784-787, 1975
INTRODUCTION INCE the published evaluation of an avirulent Pasteurella multocida drinking water vaccine by Bierer and Derieux (1972), this type vaccine has been used rather extensively by commercial turkey producers in the Carolinas (Derieux, 1974) and is referred to as the C.U. (Clemson University) strain drinking water vaccine. The usual procedure in the field has been to administer an injectable oil base P. multocida bacterin to each turkey at 8 weeks of age and then to give the drinking water vaccine three to four weeks later. The Clemson Livestock Laboratory at Columbia, South Carolina reported 5 outbreaks of fowl cholera in turkeys during 1973 alone that occurred around 5 to 6 weeks of age
and prior to the initiation of a fowl cholera control program at 8 weeks of age. In view of this, an experiment was conducted to obtain information relative to the immunologic response of small turkey poults to an avirulent P. multocida vaccine administered in the drinking water.
S
*Published with approval of the Director as Technical Contribution No. 1204.
MATERIALS AND METHODS One hundred and sixty baby turkey poults were obtained from a commercial turkey hatchery. These poults had not been injected with any type of medication or vaccine and had not been sexed. According to hatchery records the poults were the progeny of a breeder flock that had been on a fowl cholera control program that included the use of the C.U. strain drinking water vaccine prior to the time the breeders came into production. The vaccine strain has been described in a previous paper (Bierer and Derieux, 1972). Twenty of the day old poults were placed
784
Downloaded from http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
ABSTRACT Fowl cholera has been reported in turkey poults in South Carolina between 5 and 6 weeks of age. An experiment was conducted to determine the earliest age that poults could be vaccinated with the Clemson University Pasteurella multocida strain (using the drinking water route) that would result in an effective immunity at 5 to 6 weeks of age. In groups of vaccinates challenged with a virulent P. multocida (P-1059 strain) at 5 weeks of age, none were infected in groups vaccinated at either 20 days or at 25 days, 1 of 10 vaccinated at 15 days, 3 of 10 vaccinated at 10 days, 6 of 10 vaccinated at 5 days, and 8 of 10 became infected of those vaccinated at 1 day of age. Eight of 10 of each of 2 groups of nonvaccinates developed evidence of infection. In groups of vaccinated challenged at 12 weeks of age, 2 of 10 turkeys became infected that had been vaccinated at 25 days of age, 4 of 10 that had been vaccinated at 20 days, 8 of 10 vaccinated at 15 days, 8 of 10 vaccinated at 10 days, 8 of 9 vaccinated at 5 days, and 8 of 9 became infected of those vaccinated at 1 day of age. All 10 of one group of nonvaccinates and all 9 of a second group developed evidence of infection. These results suggest, under the conditions of this experiment, that turkey poults vaccinated at 35 days of age and poults vaccinated at 20 days of age experienced an excellent immunological response by 5 weeks of age. This response was less evident but, for the most part, still present, at 12 weeks of age.
PASTEURELLA
MULTOCIDA VACCINE
At 3 weeks of age, each group of poults was moved to separate growing units. 2 At 5 weeks of age, half of each of the 8 groups were placed in each of 8 individual floor pen units. The following day the poults remaining in the growing units were exposed to a highly virulent P. multocida (P-1059 strain) strain obtained from Kenneth Heddleston, Principal Research Microbiologist at the National Animal Disease Laboratory at Ames, Iowa. All 8 of the groups in floor pens were exposed to the highly virulent P. multocida strain at 12 weeks of age. Exposure, in each instance, was accomplished by the drinking water route, using a 1:50 dilution of a 20
1. Petersime Brooder Units, Model 2SD modified to single deck units. Petersime Incubator Company, Gettysburg, Ohio 45328. 2. Petersime Finishing Unit No. 4 modified to single deck units. This is a unit designed for turkey research manufactured by the Petersime Incubator Company, Gettysburg, Ohio 45328.
hours old brain-heart infusion broth culture as the only source of drinking water daily, for 3 consecutive days. Drinking water was not withheld prior to the use of this culture. Observations on all groups exposed to the highly virulent strain were discontinued 2 weeks after such exposure. Mortality data was recorded daily and the number morbid at termination was recorded. Mortality, plus morbidity was then recorded as the total number infected. The experimental facilities were housed in a well constructed old dairy barn that had been converted into areas for poultry research. The 8 floor pen units, used to house the eight treatment groups after 5 weeks of age, were of concrete block construction with a separate entrance into an anteroom. The anteroom contained feed and water facilities for the adjacent chambers and each chamber was 2.7 x 3.4 meters. These units were routinely cleaned and disinfected and the chambers prepared with fresh, clean shavings litter prior to their use. There were no other disease experiments in progress on the immediate premises. All groups were maintained in a manner that was designed to prevent direct contact of the birds between the different treatment groups. RESULTS AND DISCUSSION The results of the challenge with the highly virulent P. multocida strain at 5 weeks of age are given in Table 1. All 20 of the turkeys vaccinated at 25 and at 20 days of age survived this challenge and remained clinically healthy during the 2 weeks post challenge period of observation. Evidence of infection appeared in groups vaccinated at 15 days, 10 days, 5 days and at 1 day. The infection rate, in these groups, increased from 1 turkey in the 10 vaccinated at 15 days of age to 8 of the 10 turkeys vaccinated at 1 day of age. The results of the challenge with the virulent P. multocida strain at 12 weeks of age are given in Table 2. Fowl cholera infection
Downloaded from http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
in each of 8 brooder units. 1 C.U. strain drinking water vaccine was administered in the drinking water to the poults in the first group during the first day of age and daily thereafter, for a total of 3 consecutive days, as the only source of drinking water. The vaccine mixture was a 1:50 dilution of a 20 hours old brainheart infusion broth culture of the C.U. strain. A tenfold dilution plate count technique (Heddleston, 1972) on dextrose starch agar established that each ml. of water vaccine contained approximately 20,000,000 viable bacilli when prepared. The second group of poults received the vaccine at 5 days of age, the third at 10 days, the fourth at 15 days, the fifth at 20 days, and the sixth group of poults at 25 days of age. In each instance, as with the first group, the vaccine was given as the only source of drinking water daily, for 3 consecutive days. Drinking water was not withheld prior to the use of the vaccine. The 20 poults in each of the seventh and eighth groups were not vaccinated.
785
— —
Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Nonvaccinated Nonvaccinated
Treatment status
— —
Age when vaccinated (days) 1 5 10 15 20 25
*Number affected over number of turkeys.
Units 1 2 3 4 5 6 7 8
2 2 1 0 0 0 0 2 3 3 1 1 1 0 0 0 1 1 4 0 2 0 0 0 0 1 0
Number died (days postchallenge) 5 6 7 8 1 1 1 0 1 1 0 0 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 3 1 0 0 9 0 0 0 0 0 0 1 0
10 0 0 0 0 0 0 1 0
2 4 5 4 2 0 0 8 9
3 3 1 3 2 0 0 0 0
Number died (days postchallenge) 5 6 4 0 0 1 0 2 0 1 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 1 0
10 0 0 0 1 0 0 0 0
TABLE 2.—Postvaccination results of challenge at twelve weeks of age with a virulent P-1059
*Number affected over number of turkeys.
Unit 1 2 3 4 5 6 7 8
Treatment status Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Vaccinated Nonvaccinated Nonvaccinated
Age when vaccinated (days) 1 5 10 15 20 25
TABLE 1.—Postvaccination results of challenge at five weeks of age with a virulent P-1059
om http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
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may make them poor subjects to vaccinate, with any type of vaccine, when a lasting immunological response is desired. The fact that the poults used in the trial described in this present paper could be, for the most part, successfully vaccinated at 25 and at 20 days of age and less successfully at 15, 10, and 5 days and at 1 day of age, suggests that some or all of the detrimental factors mentioned in this paragraph may have been at play. The reader must be reminded that it was not the objective of this experiment to determine the earliest age at which an effective immunological response would occur but, rather, the earliest age at which poults could be vaccinated that would result in an effective immunity at 5 or 6 weeks of age. The results do suggest, however, that if an immunological response did result from the day old or 5 days of age vaccinations, that there was little or no evidence of this response when challenged at 35 days (5 weeks) of age. REFERENCES Bierer, B. W.,and W. T. Derieux, 1972. Immunological response in turkeys to an avirulent Pasteurella multocida vaccine in the drinking water. Poultry Sci. 51:408-416. Bierer, B. W., T. H. Eleazer and D. E. Roebuck, 1964. Sedimentation rate, packed cell volume, buffy coat value, and rectal temperature of chickens and turkeys of various ages. J. Amer. Vet. Med. Ass. 144: 727-730. Derieux, W. T., 1974. Fowl cholera, is there hope for oral vaccine? Poultry Digest, March: 126-127. Heddleston, K. L., 1972. Personal communication, January 24, 1972. Jungherr, E., and N. Terrell, 1946. Observation of the spread of Newcastle disease. Proceedings of the 50th Annual Meeting of the U.S. Livestock Sanitary Ass., p. 170. Jungherr, E., and N. Terrell, 1948. Naturally acquired passive immunity to infectious bronchitis in chicks. Amer. J. Vet. Res. 9: 201. Shoffner, R. N., 1965. Heredity and the defective in poultry, age. In: Diseases of Poultry, Biester and Schwarte Ed., Iowa State University Press, Ames, Iowa 50010, 5th Ed., page 93.
Downloaded from http://ps.oxfordjournals.org/ at University of Southern Queensland on April 28, 2015
was observed in 2 of the 10 turkeys vaccinated at 25 days of age and in 4 of the 10 turkeys vaccinated at 20 days of age. Details relative to the challenge at 5 weeks of age and the challenge at 12 weeks of age are given in Table 1 and Table 2. During the first 5 days of age there was 1 death in the group vaccinated at 1 day and at 5 days, and 1 death in a nonvaccinated group. A bacteriological examination supplied evidence that these deaths were due to E. coli infection associated with oomphalitis. These were the only deaths during the course of this experiment other than those resulting from challenge with the virulent strain of P. multocida. The poults used in this trial were from parent stock that had been vaccinated at 27 weeks of age with the C.U. strain of Pasteurella multocida by the drinking water route. The transfer of passive immunity has been reported to occur from parent stock to offspring and to persist for as long as 4 weeks in Newcastle disease (Jungherr and Terrel, 1946) and with infectious bronchitis in chickens (Jungherr and Terrell, 1948), for example. Presumably, the presence of passive immunity to fowl cholera in baby poults could interfere with their successful vaccination by the drinking water route as described under materials and methods. Also, other factors may have contributed to the high infection rates resulting from the challenge applied to the poults vaccinated at 1 day and at 5 days of age (See Tables 1 and 2). Shoffner (1965) states that, "the embryonic chick and newly hatched chick are immunologically incompetent." Presumably, this same line of reasoning can be applied to the embryonic and newly hatched poult. Bierer et al. (1964) found that such values as sedimentation rate, erythrocyte volume, buffy coat and body temperatures were not stabilized in chicks and young turkeys until the tenth to fourteenth day of life. Thus, the physiological instability of baby chicks and baby poults
