OBSERVATIONS ON THE BRUCELLA ABORTUS INFECTION IN THE BOVINE DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By NELSON BYRON KING, B. SC., D. V. M., M. S. The Ohio State University 1957 Approved bys D- Adyiser Department of Veterinary Medicine ACKNOWLEDGMENTS This opportunity is being taken to acknowledge the helpful assistance received from many persons in this endeavor. To Director L. L. Ruramell, and Associate Director W. E. Krauss, I am indebted for the permission to under­ take these studies, and for providing the facilities and funds to carry on this work. I' am particularly indebted to Dr. W. G. Venzke who has proven to be an outstanding, sound and helpful adviser throughout the period of my enrollment in the Graduate School of the Ohio State University. To Dr. B. H. Edgington, Chairman Emeritus, of the Department of Veterinary Science, sincere thanks are due to his unselfish cooperation, and generous assistance. I am very much indebted to Dr. W. D. pounden, present Chair­ man of the Department of Veterinary Science, for the assistance received from him in furthering these investi­ gations. Assistance for which I am indeed grateful on technical problems was received from Miss Norma Prank and Dr. Walter Prajola. Mr. John Bishop, Mr. John Detweiler, Mr. Prank McDaniel, Mr. Virgil Trimmer and other members of the Veterinary Science Department have been of great assistance ii iii in helping to collect specimens, in talcing cane of the animals, and attending to numerous details associated with the work. I am also grateful to Dr. Russell Conrad for his assistance in computing the statistical analyses and to others in the Dairy Science Department for their coopera­ tive efforts in making this work possible. For assistance in preparation of charts and electrophorograms, I am in­ debted to Mr. Clark Robey. To the taxpayers of Ohio who have provided the excellent faculty and facilities for carrying on the teaching and research, I am indeed grateful. TABLE OP CONTENTS Page INTRODUCTION...................... 1 REVIEW OP THE LITERATURE............................... 3 General Information..................... 3 Definition....................................... 3 History.......................................... 3 Infection in the Cow.................... 5 Infection in the Bull............................ 6 Infection in Animals Other Than Cattle.......... 7 Susceptibility to Infection...................... 9 Public Health Aspects of Brucellosis............ 10 Distribution.................................... 12 Transmission.......................... 13 Resistance of the Organism Outside the Animal Body If? Natural Course of the Disease...................... 17 Period of Incubation............................... 20 Clinical Signs..................................... 21 Tissue Changes................................ 211- Diagnosis. .................................... 25 Tube Agglutination Test......................... 25 Rapid or Plate Agglutination Test............... 28 Factors Influencing the Blood Serum Agglutina­ tion Test............................... 29 Agglutination Test on Milk Whey................. 33 Brucella Stained Antigen Milk Tests............. 314- Agglutination Test on Bull Semen. ........... lj.0 iv V Page Agglutination Test on Uterine Fluid........... Ij.0 Other Tests Used to Diagnose Brucellosis in Animals................... :.................. 1+0 Bacteriologic Methods.......................... ij.2 Attempts to Differentiate Vaccinal from Infection Titers............................. I4.3 Methods of Prevention............................. ij.5 Control Based on Periodic Blood Tests and Segregation of Reactors..................... L|.6 Control hy Vaccination......................... J4.8 Strain 19 Vaccine........................... f?0 Duration of Resistance.................. 55 Revaccination........... 56 Persistence of Agglutination Reactions Resulting from Calfhood Vaccination.... 57 Intracutaneous Injection of Strain 19 . Vaccine................................ 58 Huddleson's Mucoid Vaccine................. 60 Other Immunizing Agents Used to Control Bovine Brucellosis.............. 65 Chemotherapy...................................... 67 EXPERIMENTAL PROCEDURE........ 7J4. The Use of Huddleson's Brucella M Vaccine Under Field Conditions........ 7i| The Application of a Suggested Method for Differ­ entiating Vaccinal and Infection Titers in Cattle Known to Be Infected with Brucella abortus......... 87 The Use of Paper-Strip Electrophoresis as a Means for Differentiating Vaccinal and Br. abortus Infection Titers................................ 99 vi Page The Influence of Some Shipping Fever Bacterins on the Brucella Sero Agglutinin Titer of Cattle........................................... 121 The Survival of Br. abortus, U.S.D.A. Strain . 2308, in Manure Pit Studies..................... 132 The Treatment of Brucella abortus Infection in the Bovine ..... ....;.................... 1)])) GENERAL SUMMARY ANDCONCLUSIONS ...................... l£5 REFERENCES........................................... l£8 LIST OP TABLES , . _ _ Page Table 1. Blood Test Negative Animals Injected with "M" Vaccine at the Beginning of the Test.................... : .......... 78 Table 2. The Classification of Brucella M Vac- . cinated Animals at the Termination of the Test.................. 79 Table 3» Calvings During the Test Period............ 80 Table II. Agglutination Negative Nonvaccinated Controls................................. 82 Table £. A Comparison of the Brucella M Vaccinates and the Controls at the Termination of the Experiment.......... 83 Table 6. Changes in Agglutination Titer Following the Injection of Infected Cattle with Brucella abortus Strain 19 Vaccine...... 90 Table 7* The Application of the Differential Test on Cattle Experimentally Infected with Brucella abortus......................... 91 Table 8. Fluctuations of Agglutinin Titers of Nonvaccinated Cattle Experimentally In­ fected with Brucella abortus. .......... 93 Table 9- Fluctuations of Agglutinin Titers of TJn- exposed Cattle Vaccinated Between Seven and Twelve Months of Age with Strain 19 Vaccine............... 9i|. Table 10. The Percentage Distribution of the Globulin Fractions of the Serums of the Calves Two Meeks Following Vaccination with Strain 19 Vaccine. (Determined by Paper-Strip Electrophoresis).......................... ll£ Table 11. The Percentage Distribution of the Globulin Fractions of the Serums of Adult Cattle Two Weeks Following Vaccination with Strain 19 Vaccine. (Determined by Paper- Strip Electrophoresis).................... 115 Table 12. The Percentage Distribution of the Globulin Fractions of the Adult Infected Animals. (Determined by Paper-Strip Electro­ phoresis)................................. 116 vii viii Page Table 1 3 . The Effect of Some Hemorrhagic Septi­ cemia Biologicals on Brucella Sero Agglutination Titers................... 128 Table llj.. The Effect of Some Hemorrhagic Septicemia Biologicals on Brucella Sero Agglu­ tination Titers......................... 129 Table l£. The Effect of Sulfadiazine and Blood Plasma Transfusions on Brucella abortus Infec­ tion in Dairy Cattle................... II4.7 Table 16. The Effect of Sulfadiazine and Colmetanese on Brucella abortus Infection.......... llj.8 Table 17. The Effect of Sulfadiazine and Strepto­ mycin on Brucella abortus Infection in Dairy Cattle............................ 1^0 Table 18. The Effect of Aureomycin and Streptomycin on Brucella abortus Infection in Dairy Cattle.................................. 1^1 LIST OP ILLUSTRATIONS Number Page 1. A Photograph of the Variable Voltage Regu­ lated Power Supply and the Experimental Horizontal Cell Used in Electrophoretic Studies..................................... 105 2. A Photograph of the Recording Photometer Used in the Electrophoretic Studies......... 108 3. A.Photograph of an Electrophorogram Showing the Pour Main Components of Bovine Serum.... 109 I4. A Photograph of a Stained Filter Paper Show­ ing the Pour Main Components of Bovine Serum......................... 110 5. A Photograph of Two Electrophorograms Showing the Distribution of Serum Proteins of a Calf ,(1187) 2lj. Hours after Ingestion of Colostrum and at One Month of Age.......... Ill 6 . A Photograph of Two Electrophorograms Showing the Distribution of Serum Proteins of Colostrum-Deprived Calf (1185) 2I4 Hours after Birth and One Month of Age........... 112 7. A Photograph of the Electrophorograms Showing the Distribution of the Serum Proteins of the Colostrum-Fed (1187) and Colostrum- Deprived Calves (1185) at Pour Months of Age 113 8 . A Photograph of the Electrophorograms Showing the Distribution of the Serum Proteins Prior to and Two Weeks after Vaccination with Strain 19 Vaccine. (Cow 14.0 8 )........... 122 9. A Photograph of the Electrophorograms Showing the Distribution of the Serum Proteins Prior to, and Two Weeks after Vaccination with Strain 19 Vaccine (Cow [426............ 123 10. A Photograph of the Electrophorograms Showing the Distribution of the Serum Proteins Prior to, and 2|r Months after Brucella Infection................... ............... 12I4. 11. A Photograph Showing the Construction of the Manure Pit..................... 1314. ix Number 12. A Photograph Showing the Doors on the Top of the Manure Pit..................... 13. Manure Pit and Mean Weather Temperatures for a Summer Observation Period....... ll^. Manure Pit and Mean