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TOXIC and THERAPEUTIC EVALUATION OP STILBAMIDINE and HYDROXYSTILBAMIDINE in CANINE BLASTOMYCOSIS DISSERTATION Presented in Parti

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TOXIC and THERAPEUTIC EVALUATION OP STILBAMIDINE and HYDROXYSTILBAMIDINE in CANINE BLASTOMYCOSIS DISSERTATION Presented in Parti TOXIC AND THERAPEUTIC EVALUATION OP STILBAMIDINE AND HYDROXYSTILBAMIDINE IN CANINE BLASTOMYCOSIS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy In the Graduate School of The Ohio State University By ANDRE""LAGACE', D.V.M. The Ohio State University 1957 Approved by: 7?. Adviser Department of Veterinary Pathology ACKN OWLEDGMESTTS To Dr* C.R. Cole, chairman of the Department of Veteri­ nary Pathology at Ohio State University, I express my most sincere gratitude. Without his guidance, encouragement and understanding, it would have been impossible to ac­ complish this work. To the Quebec Agricultural Research Council, I am very grateful for their generous scholarship. To Dr G. Labelle, dean of the Quebec School of Veteri­ nary Medicine, I extend my thanks. To the techrJ cal personnel of the department I am very grateful. To my wife, Jeannine, for her encouragement, comfort and understanding during those years of studies, I will never be able to put into word3 all my appreciation. ii TABLE OP CONTENTS Page INTRODUCTION AND OBJECTIVES ..................... 1 LITERATURE R E V I E W ............................... 1|. Stllbamidine and hydroxystllbamidine Physical and chemical p r o p e r t i e s ............ k Pharmacologic action and toxicity .......... 7 Other pharmacologic effects .......... 2J+ Mechanism of action ....................... 2ij. Fungistatic properties ............... 27 Treatment of blastomycosis................... 30 TOXICITY OF STILBAMIDINE AND HYDROXYSTILBAMIDINE FOR D O G S ..................................... 33 Objectives .................. 33 Materials and Methods ....................... 33 Results ........................ 35 A) Pilot injection of stllbamidine ..... 36 B) Effects of high doses of stllbamidine . 1+3 C) Effects of low doses of stllbamidine . 53 D) Effects of high doses of hydroxystil- b a m i d i n e .......................... 66 E) Effects of low doses of hydroxystil- b a m i d l n e ............................... 71+ F) Schedule modification and therapeutic trial using dosage of dlamldlnes .... 79 G) Stllbamidine and hydroxystllbamidine for the treatment of canine blastomycosis • • 67 THE PATHOGENESIS OF STILBAMIDINE AND HYDROXYSTIL­ BAMIDINE TOXICITY ............ 91 Objectives . ........ 91 Materialsj Methods and R e s u l t s .............. 91 H) Delay in toxicity after fatal dosage of diami di n e s .......... 91 iii iv Page I) Effect of concurrent administration of corti­ sone (10 mg/kg) and diamidines.......... 98 J) Effect of histamine phosphate ...•••• 109 K) Effect of concurrent administration of anti­ histamine and diamidines ..••••.. 110 L) Comparison of thiamine deficiency in cats and dogs with diamidine toxicity in dogs • llij. EFFECTS OF STILBAMIDINE AND 2-HYDR0XYSTILBAMIDINE ON EXPERIMENTAL CANINE BLASTOMYCOSIS..........135 Objectives ....... ..................... 135 Materials and Methods 135 Results ..... Ikl M) Dogs inoculated intratracheally with 0.25 ml of packed cells ..................... li+l N) Dogs inoculated intratracheally with 0.£0 ml of packed cells ..................... lij.6 0) Dogs inoculated intravenously with 0.25 ml of packed cells ......... ....... 168 P) Dogs inoculated intravenously with 0.1 ml of packed c e l l s ......................... 181 Q) Dogs inoculated subcutaneously with 1.0 ml of packed yeast ...••••. ....... 187 DISCUSSION OF TREATMENT OF BLAST0MTC0SIS ........ 201 SUMMARY AND CONCLUSIONS ......................... 211 BIBLIOGRAPHY 220 LIST OF TABLES Table Page 1. RESUME OF ALL TEST ANIMALS FOR STILBAMIDINE TOXICITY ....................................... 1*1 2. RESUME' OF ALL TEST ANIMALS FOR HYDEOXYSTIL- BAMIDINE TOXICITY .......................... 1|2 3. EFFECTS OF HIGH DOSES OF STILBAMIDINE ........ I4 . EFFECTS OF LOW DOSES OF STILBAMIDINE.......... %. 5. EFFECTS OF HIGH AND LOW DOSES OF HYDROXYSTIL- BAMIDINE ..................................... 67 6 . SCHEDULE MODIFICATION OF A HIGH DOSAGE OF THE DIAMIDINES..................................... 80 7. STILBAMIDINE REGIMEN ......................... 8 8 8 . HYDROXYSTILBAMIDINE R E G I M E N ................... 90 9. LIST OF ALL TEST ANIMALS FOR STUDY OF PATHOGENESIS ................................. 92 10. DELAY IN TOXICITY AFTER FATAL DOSAGE OF DIAMIDINES 93 1 1 . CONCURRENT ADMINISTRATION OF CORTISONE (10 mg/kg) AND DIAMIDINES................................. 100 12. CONCURRENT ADMINISTRATION OF ANTIHISTAMINE (10 mg/ kg) AND DIAMIDINES ........................... 112 13. THIAMINE DEFICIENCY IN CATS AND DOGS ........ 120 llj.. SUMMARY OF ROUTE AND DOSE OF B^ PER MAT IT IDIS AND TREATMENT ............................... 1 3 9 l^. CLINICAL SUMMARY OF DOGS INFECTED I.T. WITH 0.5 ML YEAST ..................................... II4.7 16. CLINICAL SUMMARY OF PRETREATED DOGS INFECTED I.T. WITH 0.5 ML Y E A S T ............................. 156 17. COMPARISON OF UNTREATED, STILBAMIDINE AND HYDROXY- STILBAMIDINE TREATED DOGS INFECTED I.T. WITH 0.5 ML YEAST ..................................... 162+ vi Table Page 18. CLINICAL SUMMARY OF DOGS INFECTED I.V. WITH 0.25 ML YEAST ......................... 169 19. COMPARISON OF UNTREATED, STILBAMIDINE AND HYDROXY- STILBAMIDINE TREATED DOGS INFECTED I.V. WITH 0.25 ML Y E A S T ............................... 175 20. DISSEMINATION OF B. DERMATITIDIS IN DOGS INFECTED INTRATRACHEALLY ........................... 179 21. CLINICAL SUMMARY OF DOGS INFECTED I.V. WITH 0.10 ML Y E A S T ................................... 182 22. COMPARISON OF TREATED AND UNTREATED DOGS INFECTED 1.V. WITH 0.10 ML Y E A S T ..................... 181+ 23. DISSEMINATION OF B^ DERMATITIDIS IN DOGS INFECTED INTRAVENOUSLY OR SUBCUTANEOUSLY.............. 192 21+. DAYS BErWEEN INFECTION AND D E A T H ........... 191+ 2$, INFLUENCE OF ROUTE OF INOCULATION AND TREATMENT ON DISSEMINATION OF BLASTOMYCOSIS .......... 195 26. DISTRIBUTION OF B. DERMATITIDIS IN TREATED AND UNTREATED D O G S ............................. 196 27. DISTRIBUTION OF B. DERMATIT IDIS IN 19 TREATED AND 10 UNTREATED D O G S ....................... 197 28. DISTRIBUTION OF B. DERMATITIDIS IN 23 DOGS WITH DISSEMINATED FATAL INFECTION ................ 199 29. DISTRIBUTION OF B. DERMATITIDIS SHOWING FUNGIS­ TATIC ACTIVITY AND INCREASED OCULAR AND CENTRAL NERVOUS DISEASE IN DIAMIDINE TREATED DOGS . 200 LIST OP ILLUSTRATIONS Figure Pag© 1. PHOTOGRAPH- OPISTHOTONOS AND EXTENSOR RIGIDITY.... 40 2. PHOTOMICROGRAPH- PERIVASCULAR AND PARENCHYMAL HEMORRHAGE IN THE SUPERIOR COLLICULUS............ 40 3. PHOTOMICROGRAPH- PROTEINACEOUS MATERIAL DIFFUSING FROM VESSELS .. 55 4. PHOTOMICROGRAPH- NECROSIS, GITTER CELLS AND INCLUSION BODIES IN INFERIOR COLLICULUS.......... 55 5. PHOTOGRAPH- EXTENSIVE HEMORRHAGIC UNCEPHALOMALACIA OF THE COLLICULI.................................. 59 6 . PHOTOGRAPH- HEMORRHAGIC UNCEPHALOMALACIA OF THE COLLICULI............................ 69 7. PHOTOGRAPH- HEMORRHAGE IN CEREBELLUM AND PONS 85 8 . PHOTOMICROGRAPH- VACUOLATION AND HEMORRHAGE IN CEREBELLUM AND PONS.. ........................... 85 9. PHOTOGRAPH- TRYPAN BLUE STAINING OF CEREBELLUM AND COLLICULI............................... 101 10. PHOTOMICROGRAPH- NECROSIS OF A VESSEL............ 101 11. PHOTOGRAPH- BILATERAL HEMORRHAGIC LESIONS IN COLLICULI, CEREBELLUM AND PONS................... 103 12. PHOTOGRAPH- ENCEPHALOMALACIA IN THALAMUS......... 103 13. PHOTOMICROGRAPH- VACUOLATION AND SWOLLEN VASCULAR ENDOTHELIUM IN A CAT.............................. 122 14. PHOTOGRAPH- HEMORRHAGIC LESIONS IN CEREBELLUM AND COLLICULI..................................... 128 15. PHOTOGRAPH- HEMORRHAGIC LESIONS IN MEDULLA....... 128 16. CHART- LOCATION OF LESIONS, HIGH DOSAGE OF STILBAMIDINE...................................... 133 17. CHART- LOCATION OF LESIONS, LOW DOSAGE OF STILBAMIDINE...................................... 135 vii VIII Figure Page 18. CHART- LOCATION OF LESIONS, HIGH DOSAGE OF HYDROXY ST ILB AMID IN E ............................ 134 19. CHART- LOCATION OF LESIONS, LOW DOSAGE OF HYDROXYST ILBAMIDINE............................ 134 20. PHOTOGRAPH- LUNGS OF AN INTRATRACHEALLY INFECTED DOG................ 154 21. PHOTOMICROGRAPH- BRAIN GRANULOMA, CONTAINING GIANT CELLS AND YEAST.......................... 154 22. PHOTOMICROGRAPH- GRANULOMA IN CHOROID.......... 161 23. PHOTOMICROGRAPH- GRANULOMATOUS INFLAMMATION IN LATERAL VENTRICLE............................... 161 24. PHOTOMICROGRAPH- SUBCUTANEOUS GRANULOMA, HIGH MAGNIFICATION . ............. 172 25. PHOTOMICROGRAPH- SUBCUTANEOUS GRANULOMA........ 172 INTRODUCTION AND OBJECTIVES Recently, North American blastomycosis has been rec­ ognized more frequently in the canine specie. In the midwest area, as a systemic fungus infection it is second in impor­ tance only to histoplasmosis. Previous work done at this Uni­ versity has provided new information concerning the clinico- pathologic, radiologic, pathologic, and mycologic nature of canine North American bl&stomycoBls. The use of these diag­ nostic criteria will help the practicing veterinarian to rec­ ognize clinical cases. Now that fairly useful diagnostic procedures have been developed the urgent need is for a spe­ cific therapeutic agent. A search of the literature revealed that veterinarians have consistently failed in attempts to treat canine blasto­ mycosis. Sulfamides, penicillin, streptomycin, aureomycin, and other antibiotics, have been useless. Although stilbami- dlne has been known to possess fungistatic properties since 19<L|5 and therapeutic value in human xastomycosis since 19511
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