Selection And Clinical Trial Of The Treatment Pharmaceuticals For Canine Dermatomycosis

Canine dermatophytosis is a common zoonosis that frequently occurs in animal hospital.It has seriously affected the animal health as well as human beings together. Antifungal medicine were the most commonly chosen for the clinical therapy. However,pharmaceuticals used in the current therapy were expensive and there existed significant side-effects.Recently,with the rapid rise of resistance of fungi and the high relapse,scientists pursuit to look for a new type of antifungal drugs with broad spectrum,high efficient and low cost.Traditional Chinese medicine(TCM) is one kind of traditional advantage resources in our country,and its function and security have been certificated for centuries.The research on antifungal drug using TCM has been made many progresses.In present study,thirty-one TCM and representative strains isolated from patients with canine dermatophytosis were chosen to study the antifungal effect of single TCM or complex TCM so as to develop a new type of antifungal recipe.This study includes the following experiments.Experiment 1:Isolation,culture and characterization of pathogenic fungusThe purpose of this experiment is to isolate a few common fungi caused canine dermatomycosis.The sample were collected from suffering dogs that had been diagnosed,and they were cultured on Sabourand medium for 7～14 days.Then the fungi should be examined by microscope.The identification is made by observation of the colony surface status synchronously.Finally,four kinds of fungus are isolated: Microsporum canis,Trichophyton mentagropytes,Trichophyton rubrum and Malassezia furfur.Experiment 2:Development of antifungal medicine combining traditional Chinese medicine with chemical medicineFirstly,31 TCM with antifungal effect were selected.The tube disk approach was adopted to screen out 12 effective antifungal herbs.Secondly,the MIC(minimal inhibitory concentration) of the 12 TCM in the same method was mensurated.Three kinds of antifungal chemical medicine were selected and examined for their MIC respectively.Two Chinese herbal compounds,and selected orthogonal test(three factors and three levels) were designed to make sure the optimal combination.Results showed that an optimal complex recipe contained Radix Scutellariae,Figwort Root, Rhizoma Chuanxiong,Cortex Magnoliae Officinalis,Pseudolaricis(TCM:50 mg/mL) and Acetic Acid(1.25 mg/mL) was developed and named as "Compound Antifungal Reagents(CAR)".Experiment 3:Experiment on the stimulation and security of self-made "Compound antifungal reagents"The CAR wes daubed on naked skin of rabbits with 3 times a day,5 mL each time for 7 days continuously,the stimulation reaction of CAR to health skin and damaged skin were observed.Kun Ming mice then were orally administrated with CAR(amount to 36000 mg/kgW TCM and 3600 mg/kgW Acetic Acid) in order to test maximum tolerable daily(MTD).Results showed that "Compound antifungal reagents" had no skin stimulation or avirulence.It is safe for skin use.Experiment 4:Experiment on the cure effect of CARFour dogs were randomly assigned to one therapeutic group(three dogs) and one contrast group(one dog).Hair samples from eight parts of each dog:head,mouth,ear, back,abdomen,forelimb,posterior limb and tail were collected for the isolation and characterization of fungi.Dogs of therapeutic group were swabing down with CAR three times a week,30 minutes each time for 14 d continuously.At the same time,we daubed CAR on their skin two times a day,5 mL each time for 14 days continuously. The dog in the control was only given common bath without CAR,resulting in 22 strains of fungus isolated from 24 samples(91.7%) in the therapeutic group and 8 strains of fungus isolated in 8 samples(100%) in the control before treatment.After treatment,2 fungus in 24 samples were isolated(8.3%).The rate of bacteriostasis is 90.9%in the therapeutic group and 25%in the contrast group.