The Safety Testing Of Hydrochloride Terbinafine And The Susceptibility And Clinical Efficacy Testing Of Antifungal Agents

Terbinafine has been widely applied in the treatment of fungal skin disease on pet clinic. And its security and clinical efficacy are increasingly concerned by the majority of veterinarian and the owners. The test is about the safety testing and in vitro susceptibility testing of hydrochloride terbinafine, and the clinical efficacy testing of compound ketoconazole spray and ointment. It is aimed to provide guidance on how to apply antifungal rationally on small animal clinic.Experiment I The safety testing of hydrochloride terbinafineThe purpose of this study is to evaluate the safety of hydrochloride terbinafine capsule for dog. It was based on1425th bulletin "guidelines of safety test of pet drugs to target animal "proposed by the Ministry of Agriculture. Twenty four healthy adult dogs were selected and randomLy divided into4groups of6. Four groups were setted:the blank control group (0mg·kg-1), recommended clinical dose group (10mg·kg-1),3times the recommended clinical dose group (30mg·kg-1),5times the recommended clinical dose group (50mg·kg-1). Administered once daily for consecutive35days. Clinical symptoms were observated. Hematological and blood biochemical parameters and pathological changes were compared on the day before the test, lth week to5th after administered and2nd week after discontinuation. The result shows that as dose increased and administration time prolonged, chronic inflammation and liver and kidney damage produced by hydrochloric terbinafine tends to be more severe. It is also found that all the indicators of recommended clinical dose group were within the normal range fluctuation. And the vast majority of indicators compared with the blank control group differences were not significant (p>0.05). It proved that the recommended clinical dose (10mg·kg-1)35days of terbinafine hydrochloride for dogs is safe. Experiment Ⅱ In vitro susceptibility testing of hydrochloride terbinafine and compound ketoconazole sprayThe purpose of this study is to eluvate in vitro antifungal susceptibility of hydrochloride terbinafine capsule and compound ketoconazole spray and to guide veterinarians applying antifungal drug for pets fungal skin disease more scientific and rational. The experiment was basis on "Updated Standard for Broth Dilution Antifungal Susceptibility Testing of Yeasts (M27-A3)" and "Updated Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi (M38-A2)" proposed by the Clinical and Laboratory Standards Institute/National Committee for Clinical Laboratory Standards (CLSI/NCCLS). Microdilution method was taken. Nine common species of skin fungi were slected. They concluded5dermatophytes of T. rubrum, M. canis, M. gypseum, T. mentagrophytes, E. floccosum, and3aspergillus of A. niger, A.fumigatus, A.flavus, bisides, C. albicans. Judged the MIC according to the visual observation of broth turbidity. It shows that the MIC of hydrochloride terbinafine of nine species of fungi were0.0097,0.0064,0.0081,0.0206,0.0122,0.2000,0.2444,0.1083,18.8444μg·mL-1, the MIC of compound ketoconazole were0.1042,0.1333,0.2361,0.0868,0.1042,1.3889,0.6389,0.3194,0.2639μg·mL-1. References reported at home and abroad are basically the same.The test results reveal that hydrochloride terbinafine and compound ketoconazole both have a significant inhibitory effect on fungi skin disease of pets. The sensitivity of dermatophytes and aspergillus of hydrochloride terbinafine was higher than that of compound ketoconazole. And Candida albicans was more sensitive to compound ketoconazole than hydrochloride terbinafine.Experiment III The clinical efficacy testing of compound ketoconazole spray and ointmentThis study is undertaken to compare the clinical curative effect of the treatment on fungal skin disease of two drug formulations (spray and ointment) of compound ketoconazole. It was based on guidelines " Technical specification of experimental clinical trials" proposed by the Ministry of Agriculture. Sixty healthy adult dogs were selected. Four groups were setted:the blank control group (n=6), the drug test group (n=18), the drug control group (n=18) and the negative control group (n=18). Three most common fungal on veterinary clinic were selected:M. canis, M. gypseum, T. mentagrophytes. Dogs were infected to build fungal skin disease model and were treated with compound ketoconazole spray and ointment2weeks respectively. Clinical symptoms observations and mycological examination were taken.The result shows that the treatment efficiency of compound ketoconazole spray and ointment was88.89%and77.78%respectively. And the signs points dropped score of clinical symptoms was91.39%and89.47%respectively. It was found that two formulations for dog fungal skin diseases have a good curative effect. Overall spray was better than ointment. Compared treatment efficiency and signs points dropped score of clinical symptoms of the three fungal groups, it reveals that ointment was more effective than spray when dogs were infected by M. canis. And spray was more effective when it concerned to M. gypseum, T. mentagrophytes.