TEACHING M E A T BINDING
of heat initiated binding of poultry meat. Food Technol. 24: 42-55. Vadehra, D. V., P. G. Schnell and R. C. Baker, 1970. Effect of enzymes, muscle proteins and temperature
327
on the binding ability of poultry meat. Poultry Sci. 49: 1447. Weiss, G. H., 1971. Poultry Processing. Noyes Data Corp., Park Ridge, N.J.
An Evaluation of the Effect of Various Concentrations of the Clemson University Pasteurella multocida Drinking Water Vaccine on the Immune Response Against Fowl Cholera Disease in Turkeys* B. W.
BIERER
(Received for publication July 20. 1976)
ABSTRACT Groups of turkeys were vaccinated with various dilutions of a 24-hour-old broth culture of the Clemson University Pasteurella multocida drinking water vaccine. Corresponding numbers of viable bacilli for each vaccine concentration were established. The average total number of viable bacilli consumed by each turkey was established. Two weeks post vaccination, the immunity of the vaccinated and non vaccinated groups was challenged with a virulent P-1059 strain of P. multocida. Turkeys not vaccinated experienced a 95% infection rate when challenged. Groups of turkeys vaccinated with approximately 11 million and 23 million viable bacilli experienced severe losses (60 and 80% respectively) when challenged. Four separate groups of turkeys vaccinated with 113 million to some 2 billion viable bacilli for each bird experienced a favorable immune response and each group of turkeys compared favorably with each of the other groups. The 113 million total number of viable bacilli consumed per average turkey was the lowest concentration of bacilli resulting in a favorable immune response. Vaccination of this group of turkeys was accomplished with a drinking water dilution of 1 million viable bacilli per ml. of prepared vaccine administered over a 2 hour period once only and after a 3 hour period of water deprivation; while this procedure resulted in a favorable immune response, it was felt that the viable bacilli concentration in the drinking water (1 million viable bacilli per ml. of prepared vaccine) was probably border-line and that a much higher number of viable bacilli would probably be needed under crowded commercial turkey production field conditions. Two additional groups were vaccinated 2 hours for each of 2 consecutive days. Challenge results suggest that there was no advantage to 2 days vaccination compared to a single 1 day 2 hour vaccination period, under the conditions of this experiment. Presumably, a repeat vaccination period would be indicated under field conditions where all of the turkeys were unable to consume an adequate amount of vaccine during a single vaccination period. POULTRY SCIENCE 56: 327-330, 1977
INTRODUCTION
T
HE Clemson University (C. U.) strain of Pasteurella multocida has been and is presently being used rather extensively in the field being produced by state licensed facilities in various states (Bierer and Derieux, 1976). A federal license has been granted to a commercial company located at Gainesville, Georgia and production of a 'Published with approval of the Director as Technical Contribution No. 1385.
lyophilized product, using the C. U. strain as a drinking water vaccine against fowl cholera disease has occurred. It is the purpose of this present report to evaluate various concentrations of the C. U. strain viable bacilli administered as a drinking water vaccine against fowl cholera disease in turkeys. MATERIALS AND METHODS Ten 6-week-old turkeys were placed in each of 11 separate research units. The birds were 7 weeks old when vaccinated. The basic
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
South Carolina Agricultural Experiment Station, Clemson University, Clemson, South Carolina 29631
328
B. W. BIERER
Previous results of a tenfold dilution plate count technique (Heddleston, 1972) on dextrose starch agar plates has established that each ml. of broth culture contains approximately 1 billion viable bacilli (Bierer and Derieux, 1973; Heddleston etal., 1975). Using these previous results as a basis, the approximate numbers of viable bacilli administered per ml. of prepared vaccine ranged from 20 million in the first unit to 200,000 in the sixth. Further details along this line are given under "Results and Discussion." Four thousand ml. of prepared vaccine was placed in the waterer for each group of vaccinated turkeys and at the end of the 2 hour vaccination period unconsumed vaccine was measured for each unit so treated to determine the amount of vaccine consumed. The turkeys in each of units 7 and 8 received the 1:50 dilution of vaccine for 2 hours for each of 2 days in the same manner as the groups of vaccinated turkeys described in the foregoing. Turkeys in the ninth, tenth and eleventh units were not vaccinated.
Fourteen days after applying the various treatments described, all turkeys in each of units 1 to 10, inclusive, were exposed to a highly virulent P. multocida P-1059 strain in a culture broth-water 1:50 dilution as drinking water for 3 consecutive days. A fresh brainheart-infusion broth culture of this challenge strain was prepared each of these 3 days and given as the only source of drinking water during this period. The virulent P-1059 strain referred to was originally obtained from K. L. Heddleston, Senior Research Microbiologist at the National Animal Disease Laboratory at Ames, Iowa. The experiment was terminated 14 days after exposure to the virulent culture. At that time the number of turkeys that died in each unit was added to the number of survivors that were morbid, if any, to arrive at a total value for the turkeys in each of the treatment groups. Turkeys in the eleventh unit were neither vaccinated nor challenged and were maintained as a monitor group. The research units used were ordinary cement floor pens with shavings and each unit provided approximately 5 square feet for each turkey. Feed and water could be supplied without entering the units. All groups of turkeys were observed daily and the number of dead and morbid turkeys, if present, were recorded. The turkeys used were obtained from a commercial flock with no history of a disease problem. There were no other infectious disease experiments in progress in the research facility used during the time the work described was conducted. All units were cleaned and disinfected after each experiment and provided with fresh, clean shavings when needed. The possible ill effects, if any, of the vaccination procedure described on body weights was not determined. RESULTS AND DISCUSSION Prepared vaccine consumed ranged from 1202 ml. in the first group of turkeys to 991
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
vaccine material was a 24 hours old brainheart-infusion broth culture of the C. U. strain of P. multocida. Turkeys in the first 6 units, inclusive, received vaccine in the drinking water for 2 hours. Regular drinking water had previously been removed for a period of 3 hours and an amount of fresh feed had been placed in all units during this no water period. Turkeys in the first unit received a 1:50 dilution of the broth culture as vaccine during the vaccination period. Those in the second unit received a 1:100 dilution, in the third a 1:500 dilution, in the fourth unit a 1:1000 dilution, in the fifth a 1:5000 dilution and the turkeys in the sixth unit a 1:10,000 dilution of the broth culture as vaccine. In preparing these dilutions, the lowest dilution (1:10,000) was prepared first, the next to lowest second, etc., until all dilutions were prepared. Pipettes used in measuring were of the disposable variety and each pipette was discarded when used only once.
329
FOWL CHOLERA DISEASE IN TURKEYS
ml. in the sixth vaccinated group for a total of 6796 ml. consumed by the 60 turkeys in all six units or an average of 113 ml. of vaccine consumed per turkey, during the 2 hour vaccination period. There was no evidence of a decreased vaccine consumption due to a palatability problem with the more concentrated dilutions. A break down giving the approximate numbers of viable bacilli consumed for the average turkey in each of the 6 groups receiving the various dilutions of vaccine is given in Table 1.
TABLE 1. -Approximate numbers of viable bacilli consumed per average turkey for the various vaccine
concentrations administered and final challenge results
Research unit
Broth-water mixture ratio*
Approx. number viable bacilli per ml. vaccine
Approx. total number viable bacilli consumed per 1 average turkey*"
1 2 3 4 5 6
1:50 1:100 1:500 1:1000 1:5000 1:10,000
20 million 10 million 2 million 1 million 200,000 100,000
2,260,000,000 1,130,000,000 226,000,000 113,000,000 22,600,000 11,300,000
Mortality plus morbidity total (%) 0 20 20 10 60 80
* Broth refers to a 24-hour-old brain-heart-infusion broth culture of the C.U. strain of Pasteurella multocida administered to the 10 turkeys of each unit. ** Average vaccine consumption per turkey was 113 ml. during the 2 hour vaccination period. TABLE 2.—Detailed postvaccination results following challenge with a virulent P-1059 strain of Pasteurella multocida
Research unit 1 2 3 4 5 6 7 8 9 10 11
Broth-water mixture ratio* 1:50 1:100 1:500 1:1000 1:5000 1:10,000 1:50 (2 days) 1:50 (2 days) none none none**
1 0 0 0 0 0 0 0 0 0 0 0
2 0 0 0 0 2 0 0 0 0 3 0
Daily record of deaths 7 6 5 4 3 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 0 0 0 0 0 1 0 1 2 1 0 1 0 6 1 0 0 1 0 0 0 0 0 1 0 2 2 2 1 0 0 3 0 4 0 0 0 0 0
8 0 0 0 0 0 0 0 0 1 0 0
9-14 0 0 0 0 0 0 0 0 0 0 0
Total number died 0 2 1 0 6 8 2 1 8 10 0
Total number morbid 0 0 1 1 0 0 0 0 1 0 0
Mortality plus morbidity total
(%) 0 20 20 10 60 80 20 10 90 100 0
*Supplied as the only source of drinking water for 2 hours after water deprivation for 3 hours. Broth refers to a 24-hour-old brain-heart-infusion culture of the C.U. strain. **This group was neither vaccinated nor challenged but served as a monitor group.
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
Mortality and morbidity results, after challenge with the highly virulent P. multocida strain was 95% for the 2 groups that were
not vaccinated and ranged from 0% in the group receiving the 1:50 dilution vaccine for 2 hours only to 80% in the group receiving the 1:10,000 dilution vaccine. Details are given in Table 2. There was no evidence that vaccination for 2 days at the 1:50 dilution of the C. U. broth culture was superior to 2 hours during 1 day at the same dilution, in stimulating an effective immune response. Perhaps the 2 hours of vaccination on 2 consecutive days would be more advantageous when administered in the lower 1:5000 and 1:10,000 dilutions. The local immune response theory, as described by Bienenstock (1975), leads one
330
B. W.
for any vaccine under commercial conditions.
field
REFERENCES Bienenstock, J., 1975. The local immune response. Amer. J. Vet. Res. 36: 488-491. Bierer, B. W., and W. T. Derieux, 1973. Exposing turkeys by various routes to an avirulent and virulent strains of Pasteurella multocida. Poultry Sci. 52: 2290-2298. Bierer, B. W., and W. T. Derieux, 1976. The effect of sulfaquinoxaline feed medication on the immunologic response to a Pasteurelly multocida vaccine administered to turkeys via drinking water. Poultry Sci. 55: 209-212. Heddleston, K. L., 1972. Personal communication, January 24, 1972. Heddleston, K. L., P. A. Rebers and G. Wessman, 1975. Fowl cholera: Immunologic and serologic response in turkeys to live Pasteurella multocida vaccine administered in the drinking water. Poultry Sci. 54:217-221.
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
to believe that antigen concentration either initially or over a period of time will increase an immune response which, eventually would be indistinguishable in quality and amount from that obtained by parental immunization. Vaccination at the 1:1000 dilution appeared to be equally as effective as vaccination at the 1:50 dilution with reference to an effective immune response. It would appear, however, that a commercial vaccination procedure relying on the 1:1000 dilution of the 24-hourold brain-heart-infusion broth culture of the C. U. strain or a concentrated commercial product supplying only 1 million viable bacilli per ml. of prepared vaccine and supplying only 113 million viable bacilli per turkey for the vaccination period, that such products would be of border-line value. A substantial margin of safety is an important requirement
BIERER
of heat initiated binding of poultry meat. Food Technol. 24: 42-55. Vadehra, D. V., P. G. Schnell and R. C. Baker, 1970. Effect of enzymes, muscle proteins and temperature
327
on the binding ability of poultry meat. Poultry Sci. 49: 1447. Weiss, G. H., 1971. Poultry Processing. Noyes Data Corp., Park Ridge, N.J.
An Evaluation of the Effect of Various Concentrations of the Clemson University Pasteurella multocida Drinking Water Vaccine on the Immune Response Against Fowl Cholera Disease in Turkeys* B. W.
BIERER
(Received for publication July 20. 1976)
ABSTRACT Groups of turkeys were vaccinated with various dilutions of a 24-hour-old broth culture of the Clemson University Pasteurella multocida drinking water vaccine. Corresponding numbers of viable bacilli for each vaccine concentration were established. The average total number of viable bacilli consumed by each turkey was established. Two weeks post vaccination, the immunity of the vaccinated and non vaccinated groups was challenged with a virulent P-1059 strain of P. multocida. Turkeys not vaccinated experienced a 95% infection rate when challenged. Groups of turkeys vaccinated with approximately 11 million and 23 million viable bacilli experienced severe losses (60 and 80% respectively) when challenged. Four separate groups of turkeys vaccinated with 113 million to some 2 billion viable bacilli for each bird experienced a favorable immune response and each group of turkeys compared favorably with each of the other groups. The 113 million total number of viable bacilli consumed per average turkey was the lowest concentration of bacilli resulting in a favorable immune response. Vaccination of this group of turkeys was accomplished with a drinking water dilution of 1 million viable bacilli per ml. of prepared vaccine administered over a 2 hour period once only and after a 3 hour period of water deprivation; while this procedure resulted in a favorable immune response, it was felt that the viable bacilli concentration in the drinking water (1 million viable bacilli per ml. of prepared vaccine) was probably border-line and that a much higher number of viable bacilli would probably be needed under crowded commercial turkey production field conditions. Two additional groups were vaccinated 2 hours for each of 2 consecutive days. Challenge results suggest that there was no advantage to 2 days vaccination compared to a single 1 day 2 hour vaccination period, under the conditions of this experiment. Presumably, a repeat vaccination period would be indicated under field conditions where all of the turkeys were unable to consume an adequate amount of vaccine during a single vaccination period. POULTRY SCIENCE 56: 327-330, 1977
INTRODUCTION
T
HE Clemson University (C. U.) strain of Pasteurella multocida has been and is presently being used rather extensively in the field being produced by state licensed facilities in various states (Bierer and Derieux, 1976). A federal license has been granted to a commercial company located at Gainesville, Georgia and production of a 'Published with approval of the Director as Technical Contribution No. 1385.
lyophilized product, using the C. U. strain as a drinking water vaccine against fowl cholera disease has occurred. It is the purpose of this present report to evaluate various concentrations of the C. U. strain viable bacilli administered as a drinking water vaccine against fowl cholera disease in turkeys. MATERIALS AND METHODS Ten 6-week-old turkeys were placed in each of 11 separate research units. The birds were 7 weeks old when vaccinated. The basic
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
South Carolina Agricultural Experiment Station, Clemson University, Clemson, South Carolina 29631
328
B. W. BIERER
Previous results of a tenfold dilution plate count technique (Heddleston, 1972) on dextrose starch agar plates has established that each ml. of broth culture contains approximately 1 billion viable bacilli (Bierer and Derieux, 1973; Heddleston etal., 1975). Using these previous results as a basis, the approximate numbers of viable bacilli administered per ml. of prepared vaccine ranged from 20 million in the first unit to 200,000 in the sixth. Further details along this line are given under "Results and Discussion." Four thousand ml. of prepared vaccine was placed in the waterer for each group of vaccinated turkeys and at the end of the 2 hour vaccination period unconsumed vaccine was measured for each unit so treated to determine the amount of vaccine consumed. The turkeys in each of units 7 and 8 received the 1:50 dilution of vaccine for 2 hours for each of 2 days in the same manner as the groups of vaccinated turkeys described in the foregoing. Turkeys in the ninth, tenth and eleventh units were not vaccinated.
Fourteen days after applying the various treatments described, all turkeys in each of units 1 to 10, inclusive, were exposed to a highly virulent P. multocida P-1059 strain in a culture broth-water 1:50 dilution as drinking water for 3 consecutive days. A fresh brainheart-infusion broth culture of this challenge strain was prepared each of these 3 days and given as the only source of drinking water during this period. The virulent P-1059 strain referred to was originally obtained from K. L. Heddleston, Senior Research Microbiologist at the National Animal Disease Laboratory at Ames, Iowa. The experiment was terminated 14 days after exposure to the virulent culture. At that time the number of turkeys that died in each unit was added to the number of survivors that were morbid, if any, to arrive at a total value for the turkeys in each of the treatment groups. Turkeys in the eleventh unit were neither vaccinated nor challenged and were maintained as a monitor group. The research units used were ordinary cement floor pens with shavings and each unit provided approximately 5 square feet for each turkey. Feed and water could be supplied without entering the units. All groups of turkeys were observed daily and the number of dead and morbid turkeys, if present, were recorded. The turkeys used were obtained from a commercial flock with no history of a disease problem. There were no other infectious disease experiments in progress in the research facility used during the time the work described was conducted. All units were cleaned and disinfected after each experiment and provided with fresh, clean shavings when needed. The possible ill effects, if any, of the vaccination procedure described on body weights was not determined. RESULTS AND DISCUSSION Prepared vaccine consumed ranged from 1202 ml. in the first group of turkeys to 991
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
vaccine material was a 24 hours old brainheart-infusion broth culture of the C. U. strain of P. multocida. Turkeys in the first 6 units, inclusive, received vaccine in the drinking water for 2 hours. Regular drinking water had previously been removed for a period of 3 hours and an amount of fresh feed had been placed in all units during this no water period. Turkeys in the first unit received a 1:50 dilution of the broth culture as vaccine during the vaccination period. Those in the second unit received a 1:100 dilution, in the third a 1:500 dilution, in the fourth unit a 1:1000 dilution, in the fifth a 1:5000 dilution and the turkeys in the sixth unit a 1:10,000 dilution of the broth culture as vaccine. In preparing these dilutions, the lowest dilution (1:10,000) was prepared first, the next to lowest second, etc., until all dilutions were prepared. Pipettes used in measuring were of the disposable variety and each pipette was discarded when used only once.
329
FOWL CHOLERA DISEASE IN TURKEYS
ml. in the sixth vaccinated group for a total of 6796 ml. consumed by the 60 turkeys in all six units or an average of 113 ml. of vaccine consumed per turkey, during the 2 hour vaccination period. There was no evidence of a decreased vaccine consumption due to a palatability problem with the more concentrated dilutions. A break down giving the approximate numbers of viable bacilli consumed for the average turkey in each of the 6 groups receiving the various dilutions of vaccine is given in Table 1.
TABLE 1. -Approximate numbers of viable bacilli consumed per average turkey for the various vaccine
concentrations administered and final challenge results
Research unit
Broth-water mixture ratio*
Approx. number viable bacilli per ml. vaccine
Approx. total number viable bacilli consumed per 1 average turkey*"
1 2 3 4 5 6
1:50 1:100 1:500 1:1000 1:5000 1:10,000
20 million 10 million 2 million 1 million 200,000 100,000
2,260,000,000 1,130,000,000 226,000,000 113,000,000 22,600,000 11,300,000
Mortality plus morbidity total (%) 0 20 20 10 60 80
* Broth refers to a 24-hour-old brain-heart-infusion broth culture of the C.U. strain of Pasteurella multocida administered to the 10 turkeys of each unit. ** Average vaccine consumption per turkey was 113 ml. during the 2 hour vaccination period. TABLE 2.—Detailed postvaccination results following challenge with a virulent P-1059 strain of Pasteurella multocida
Research unit 1 2 3 4 5 6 7 8 9 10 11
Broth-water mixture ratio* 1:50 1:100 1:500 1:1000 1:5000 1:10,000 1:50 (2 days) 1:50 (2 days) none none none**
1 0 0 0 0 0 0 0 0 0 0 0
2 0 0 0 0 2 0 0 0 0 3 0
Daily record of deaths 7 6 5 4 3 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 0 0 0 0 0 1 0 1 2 1 0 1 0 6 1 0 0 1 0 0 0 0 0 1 0 2 2 2 1 0 0 3 0 4 0 0 0 0 0
8 0 0 0 0 0 0 0 0 1 0 0
9-14 0 0 0 0 0 0 0 0 0 0 0
Total number died 0 2 1 0 6 8 2 1 8 10 0
Total number morbid 0 0 1 1 0 0 0 0 1 0 0
Mortality plus morbidity total
(%) 0 20 20 10 60 80 20 10 90 100 0
*Supplied as the only source of drinking water for 2 hours after water deprivation for 3 hours. Broth refers to a 24-hour-old brain-heart-infusion culture of the C.U. strain. **This group was neither vaccinated nor challenged but served as a monitor group.
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
Mortality and morbidity results, after challenge with the highly virulent P. multocida strain was 95% for the 2 groups that were
not vaccinated and ranged from 0% in the group receiving the 1:50 dilution vaccine for 2 hours only to 80% in the group receiving the 1:10,000 dilution vaccine. Details are given in Table 2. There was no evidence that vaccination for 2 days at the 1:50 dilution of the C. U. broth culture was superior to 2 hours during 1 day at the same dilution, in stimulating an effective immune response. Perhaps the 2 hours of vaccination on 2 consecutive days would be more advantageous when administered in the lower 1:5000 and 1:10,000 dilutions. The local immune response theory, as described by Bienenstock (1975), leads one
330
B. W.
for any vaccine under commercial conditions.
field
REFERENCES Bienenstock, J., 1975. The local immune response. Amer. J. Vet. Res. 36: 488-491. Bierer, B. W., and W. T. Derieux, 1973. Exposing turkeys by various routes to an avirulent and virulent strains of Pasteurella multocida. Poultry Sci. 52: 2290-2298. Bierer, B. W., and W. T. Derieux, 1976. The effect of sulfaquinoxaline feed medication on the immunologic response to a Pasteurelly multocida vaccine administered to turkeys via drinking water. Poultry Sci. 55: 209-212. Heddleston, K. L., 1972. Personal communication, January 24, 1972. Heddleston, K. L., P. A. Rebers and G. Wessman, 1975. Fowl cholera: Immunologic and serologic response in turkeys to live Pasteurella multocida vaccine administered in the drinking water. Poultry Sci. 54:217-221.
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on May 2, 2015
to believe that antigen concentration either initially or over a period of time will increase an immune response which, eventually would be indistinguishable in quality and amount from that obtained by parental immunization. Vaccination at the 1:1000 dilution appeared to be equally as effective as vaccination at the 1:50 dilution with reference to an effective immune response. It would appear, however, that a commercial vaccination procedure relying on the 1:1000 dilution of the 24-hourold brain-heart-infusion broth culture of the C. U. strain or a concentrated commercial product supplying only 1 million viable bacilli per ml. of prepared vaccine and supplying only 113 million viable bacilli per turkey for the vaccination period, that such products would be of border-line value. A substantial margin of safety is an important requirement
BIERER
